Cycloalkane-1,3-diamine derivative

ABSTRACT

The present invention provides a compound or a pharmaceutically acceptable salt thereof having an inhibitory action on the interaction between menin and an MLL protein. The compound represented by the formula (1) or a pharmaceutically acceptable salt thereof.wherein, in the formula (1), the dotted circle, R1, R2, R3, R4, R5, R6, R7, R8, Ring Q1, W, m and n are each as defined in the description.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International Application No.PCT/JP2019/048834, filed Dec. 5, 2019, which claims priority to JapaneseApplication No. 2018-229397, filed Dec. 6, 2018, each expresslyincorporated herein by reference in its entirety.

TECHNICAL FIELD

The present invention relates to low-molecular compounds or apharmaceutically acceptable salt thereof that inhibit the interactionbetween menin and an MLL protein.

BACKGROUND ART

Chromosomal translocation of MLL (Mixed-Lineage Leukemia) gene isobserved in infant leukemia and some poor-prognosis leukemia cases. As aresult of chromosomal translocation, MLL fuses with 70 or more varioustranslocation partner genes at its amino-terminus to express an MLLfusion protein. Wild-type MLL constitutes a transcriptional regulatorycomplex that modifies the chromatin structure, specifically methylateslysine at the 4th position of histone H3, and plays an extremelyimportant role in the transcriptional regulation of gene cluster (e.g.,HOX gene cluster, etc.) involved in hematopoiesis and development.Meanwhile, the MLL fusion protein, whose expression is induced bychromosomal translocation, has lost the histone methylase activity, butpermanently activates gene cluster (e.g., HOX and MEIS1 genes, etc.)involved in cell differentiation control. As a result, abnormal cellgrowth and inhibition of differentiation induction of hematopoieticcells are triggered, which leads to onset of leukemia. Leukemia with MLLgene mutation has poor prognosis, and the standard treatment methodscurrently used for leukemia treatment have not been sufficientlyeffective. For this reason, development of a new treatment method isstrongly desired.

Menin is a tumor-suppressor protein identified as a causal factor ofmultiple endocrine neoplasia type 1 (MEN1), which is one of autosomaldominant hereditary tumor syndromes, and characterized by tumorigenesisin multiple endocrine organs. Menin is an ubiquitously expressednucleoprotein that interacts with a wide variety of proteins and isinvolved in various cellular processes. It is considered that thebiological functions of menin can be tumor-suppressing ortumor-promoting, and dependent on the cell context. Menin interacts withthe amide-terminus of MLL1, and functions as a carcinogenic cofactorthat increases the transcription of gene cluster such as HOX and MEIS1.It is known that the interaction between menin and an MLL fusion proteinis essential for abnormal activation of a series of gene cluster causedby the MLL fusion protein, and onset of leukemia (Non-Patent Documents 1and 2). Thus, it is expected that inhibition of the interaction betweenmenin and an MLL fusion protein contributes to the treatment and/orprophylaxis of leukemias involving chromosomal translocations of MLLgene and other leukemia/blood cancers accompanied with constantexpression of HOX and MEIS1 genes. Accordingly, for example, thecreation of a drug that inhibits the interaction between menin and anMLL fusion protein is extremely significant in terms of providing a newoption for cancer treatment.

A plurality of compounds having an inhibitory activity on theinteraction between menin and an MLL protein have been already known(Patent Documents 1 to 4, Non-Patent Documents 3 to 5).

DOCUMENT LIST Patent Document

-   Patent Document 1: WO 2017/161028 pamphlet-   Patent Document 2: WO 2018/053267 pamphlet-   Patent Document 3: WO 2018/109088 pamphlet-   Patent Document 4: WO 2018/024602 pamphlet

Non-Patent Document

-   Non-Patent Document 1: Chen et al., Proc. Natl. Acad. Sci., 2006,    103, 1018-1023.-   Non-Patent Document 2: Yokoyama et al., Cell, 2005, 123, 207-218.-   Non-Patent Document 3: Grembecka et al., Nat. Chem. Biol., 2012, 8,    277-284.-   Non-Patent Document 4: Shi et al., Blood, 2012, 120, 4461-4469.-   Non-Patent Document 5: Borkin et al., Cancer Cell, 2015, 27,    589-602.

SUMMARY OF THE INVENTION Problem to be Solved by the Invention

The present invention provides a novel low-molecular compound having aninhibitory action on the interaction between menin and an MLL protein(hereinafter, sometimes to be referred to as a menin-MLL inhibitoryaction), which is useful as a medicament for the treatment and/orprophylaxis of diseases dependent on the interaction between menin andan MLL protein.

Means of Solving the Problems

The present inventors have conducted research on novel low-molecularcompounds with the aim of developing a menin-MLL inhibitor, and havefound that a compound having a specific structure or a pharmaceuticallyacceptable salt thereof disclosed in the present invention has amenin-MLL inhibitory action, and is useful as a medicament for thetreatment and/or prophylaxis of diseases (e.g., cancer or diabetes)dependent on the interaction between menin and an MLL protein, andcompleted the present invention based on these findings. The compoundsor pharmaceutically acceptable salts thereof disclosed in the presentinvention have not been known so far, and their pharmacologicalactivities are also unknown.

The present invention relates to the following [1] to [92].

[1] A compound represented by the following general formula (1) or apharmaceutically acceptable salt thereof:

whereinthe dotted circle indicates that the ring is aromatic, R¹ and R² areeach independently a hydrogen atom or a C₁₋₆ alkyl group,one of R³ and R⁴ is a hydrogen atom, a hydroxy group, a halogen atom, aC₁₋₆ alkoxy group, a di(C₁₋₆ alkyl)carbamoyl group, or an oxazolylgroup, andthe other of R³ and R⁴ is a hydrogen atom, a hydroxy group, a halogenatom, or a C₁₋₆ alkoxy group,R⁵ is a hydrogen atom, a C₁₋₆ alkyl group, or a hydroxy C₁₋₆ alkylgroup,R⁶ is a hydrogen atom, a C₁₋₆ alkyl group, a halogen atom, a C₁₋₆ alkoxygroup, an amino group, or a C₁₋₆ alkylamino group,R⁷ and R⁸ are taken together with the carbon atom to which R⁷ is bondedand the carbon atom to which R⁸ is bonded to form any of the followingformulas (2A) to (2C):

-   -   wherein    -   the dotted circle indicates that the ring is aromatic,    -   the carbon atom marked with a is the carbon atom to which R⁸ is        bonded,    -   the carbon atom marked with b is the carbon atom to which R⁷ is        bonded,    -   X is CH or a nitrogen atom, and    -   R⁹ is a halogeno C₁₋₆ alkyl group, a C₃₋₈ cycloalkyl group, a    -   C₃₋₈ cycloalkyl C₁₋₆ alkyl group, a C₁₋₆ alkoxy C₁₋₆ alkyl        group, or an oxetanyl group, or        R⁷ is a hydrogen atom, and R⁸ is the following formula (3):

-   -   wherein    -   * indicates a bonding site,    -   R¹⁰ is a di(C₁₋₆ alkyl) carbamoyl group, a (C₁₋₆        alkyl)pyrimidinyl group, a (C₁₋₆ alkyl)phenyl group, or a (C₁₋₆        alkyl)pyrazolyl group,    -   R¹¹ is a hydrogen atom or a halogen atom, and    -   R¹² is a halogen atom,        m is 1 or 0,        n is 1 or 2,        Ring Q¹ is a 6-membered aromatic ring optionally containing one        nitrogen atom in the ring (the aromatic ring optionally has one        or two substituents independently selected from the following        Group A), a 5-membered aromatic heterocycle containing, in the        ring, one or two heteroatoms independently selected from the        group consisting of a nitrogen atom and a sulfur atom (the        aromatic heterocycle optionally has one substituent        independently selected from the following Group A), a C₃₋₈        cycloalkane ring optionally having one substituent independently        selected from the following Group A, a C₄₋₈ cycloalkene ring        optionally having one substituent independently selected from        the following Group A, a 4- to 8-membered saturated heterocycle        containing one nitrogen atom in the ring (the saturated        heterocycle optionally has one substituent independently        selected from the following Group A), or a 9-membered bicyclic        aromatic heterocycle containing one nitrogen atom in the ring        (the bicyclic aromatic heterocycle optionally has one or two        substituents independently selected from the following Group B),        and        W is the following formula (4A) or (4B):

-   -   wherein    -   * indicates a bonding site,    -   Ring Q² is a 6-membered aromatic ring optionally containing one        nitrogen atom in the ring (the aromatic ring optionally has one        to three substituents independently selected from the following        Group C), a 6-membered aromatic heterocycle containing two        nitrogen atoms in the ring (the aromatic heterocycle optionally        has one to three substituents independently selected from the        following Group C), a 5-membered aromatic heterocycle        containing, in the ring, one to three heteroatoms independently        selected from the group consisting of a nitrogen atom, an oxygen        atom and a sulfur atom (the aromatic heterocycle optionally has        one substituent independently selected from the following Group        C), a 9- or 10-membered bicyclic aromatic or partially        unsaturated heterocycle containing, in the ring, one to three        heteroatoms independently selected from the group consisting of        a nitrogen atom and an oxygen atom (the bicyclic aromatic or        partially unsaturated heterocycle optionally has one or two        substituents independently selected from the following Group D),        a 5- to 8-membered saturated heterocycle containing, in the        ring, one or two heteroatoms independently selected from the        group consisting of an oxygen atom and a nitrogen atom (the        saturated heterocycle optionally has one substituent        independently selected from the following Group E), or a C₃₋₈        cycloalkane ring optionally having one substituent independently        selected from the following Group E, Ring Q³ is a 4- to        8-membered saturated heterocycle containing one nitrogen atom or        one oxygen atom in the ring (the saturated heterocycle        optionally has one C₁₋₆ alkylsulfonyl group), or a 6-membered        aromatic ring optionally containing one nitrogen atom in the        ring (the aromatic ring optionally has one substituent        independently selected from the following Group F),    -   Y is a single bond or an oxygen atom, and    -   Z is a single bond, an oxygen atom, —NH—, —SO₂—, a C₁₋₆ alkylene        group, *—R¹³—NHC(═O)—**, *—R⁴—O—**, or *—R⁵—NH—**,    -   wherein * is bonded to Ring Q², ** is bonded to Ring Q¹, and        R¹³, R¹⁴ and R¹⁵ are each independently a C₁₋₆ alkylene group,        Group A: a halogen atom, a hydroxy group, a C₁₋₆ alkyl group, a        C₁₋₆ alkoxy group, a hydroxy C₁₋₆ alkoxy group, a        vinylsulfonylamino(C₁₋₆ alkyl)carbamoyl group, and a        prop-2-enoylamino(C₁₋₆ alkyl)carbamoyl group,        Group B: a cyano group, a C₁₋₆ alkyl group, a halogen atom, and        a C₁₋₆ alkoxy group,        Group C: a halogen atom, a C₁₋₆ alkyl group, a C₁₋₆ alkoxy        group, a C₁₋₆ alkyl(C₁₋₆ alkylsulfonyl)amino group, a cyano        group, a C₁₋₆ alkylsulfonyl group, a C₁₋₆ alkylamino group, a        di(C₁₋₆ alkyl)amino group, a halogeno C₁₋₆ alkyl group, a C₁₋₆        alkoxy C₁₋₆ alkoxy group, a halogeno C₁₋₆ alkoxy group, a C₁₋₆        alkylsulfonyl C₁₋₆ alkyl group, a di(C₁₋₆ alkyl)sulfamoyl group,        a C₁₋₆ alkylenedioxy group, a (C₁₋₆ alkyl)carbamoyl group, a        hydroxy C₁₋₆ alkyl group, a 2-C₃₋₆ alkenoylamino group, a C₁₋₆        alkyl (2-C₃₋₆ alkenoyl)amino group, a hydroxy group, an oxo        group, a (²H3)methoxy group, and a bis[(²H3)methyl]amino group,        Group D: a halogen atom, a C₁₋₆ alkyl group, and a C₁₋₆        alkylsulfonyl group,        Group E: an oxo group, a hydroxy group, and a C₁₋₆ alkoxy group,        and        Group F: a halogen atom, and a C₁₋₆ alkoxy group.        [2] The compound according to [1], or a pharmaceutically        acceptable salt thereof, wherein R¹ is a hydrogen atom or a        methyl group.        [3] The compound according to [1], or a pharmaceutically        acceptable salt thereof, wherein R¹ is a hydrogen atom.        [4] The compound according to any one of [1] to [3], or a        pharmaceutically acceptable salt thereof, wherein R² is a        hydrogen atom or a methyl group.        [5] The compound according to any one of [1] to [3], or a        pharmaceutically acceptable salt thereof, wherein R² is a        hydrogen atom.        [6] The compound according to any one of [1] to [5], or a        pharmaceutically acceptable salt thereof, wherein the moiety        represented by the following formula (5) in the formula (1) is        the following formula (5A) or (5B):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded,    -   ** is bonded to the nitrogen atom to which R⁵ is bonded,    -   R¹⁶ is a hydrogen atom, a halogen atom, a hydroxy group, a        di(C₁₋₆ alkyl)carbamoyl group, an oxazol-2-yl group, or a C₁₋₆        alkoxy group,    -   R¹⁷ is a hydrogen atom or a halogen atom, and    -   R¹⁸ is a C₁₋₆ alkoxy group.        [7] The compound according to any one of [1] to [5], or a        pharmaceutically acceptable salt thereof, wherein the moiety        represented by the following formula (5) in the formula (1) is        the following formula (6A) or (6B):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded,    -   ** is bonded to the nitrogen atom to which R⁵ is bonded, and    -   R¹⁹ is a hydrogen atom, a hydroxy group, a dimethylcarbamoyl        group, an oxazol-2-yl group, or a methoxy group.        [8] The compound according to any one of [1] to [5], or a        pharmaceutically acceptable salt thereof, wherein the moiety        represented by the following formula (5) in the formula (1) is        the following formula (7A):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded,    -   ** is bonded to the nitrogen atom to which R⁵ is bonded,    -   R²⁰ is a hydrogen atom or a hydroxy group, and    -   R²¹ is a hydrogen atom, a hydroxy group, or a C₁₋₆ alkoxy group.        [9] The compound according to any one of [1] to [5], or a        pharmaceutically acceptable salt thereof, wherein the moiety        represented by the following formula (5) in the formula (1) is        any of the following formulas (8A) to (8E):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded,    -   * * is bonded to the nitrogen atom to which R⁵ is bonded,    -   R²² is a hydrogen atom, a hydroxy group or a methoxy group,    -   R²³ is a hydroxy group or a methoxy group, and    -   R²⁴ is a hydrogen atom or a hydroxy group.        [10] The compound according to any one of [1] to [5], or a        pharmaceutically acceptable salt thereof, wherein the moiety        represented by the following formula (5) in the formula (1) is        any of the following formulas (9A) to (9C):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded, and    -   ** is bonded to the nitrogen atom to which R⁵ is bonded.        [11] The compound according to any one of [1] to [10], or a        pharmaceutically acceptable salt thereof, wherein R⁵ is a        hydrogen atom, a methyl group, an ethyl group, or a        2-hydroxyethyl group.        [12] The compound according to any one of [1] to [10], or a        pharmaceutically acceptable salt thereof, wherein R⁵ is a methyl        group.        [13] The compound according to any one of [1] to [12], or a        pharmaceutically acceptable salt thereof, wherein R⁶ is a        hydrogen atom, a methyl group, a chlorine atom, a methoxy group,        an amino group, or a methylamino group.        [14] The compound according to any one of [1] to [13], or a        pharmaceutically acceptable salt thereof, wherein R⁷ and R⁸ are        taken together with the carbon atom to which R⁷ is bonded and        the carbon atom to which R⁸ is bonded to form the following        formula (10A):

-   -   wherein    -   the dotted circle indicates that the ring is aromatic,    -   the carbon atom marked with a is the carbon atom to which R⁸ is        bonded, and    -   the carbon atom marked with b is the carbon atom to which R⁷ is        bonded.        [15] The compound according to any one of [1] to [13], or a        pharmaceutically acceptable salt thereof, wherein R⁷ is a        hydrogen atom, and R⁸ is the following formula (11A) or (11B):

-   -   wherein    -   * indicates a bonding site,    -   R²⁵ is a diisopropylcarbamoyl group, a 4-isopropylpyrimidin-5-yl        group, a 2-isopropylphenyl group, or a 1-isopropylpyrazol-5-yl        group, and    -   R²⁶ is a diisopropylcarbamoyl group.        [16] The compound according to any one of [1] to [15], or a        pharmaceutically acceptable salt thereof, wherein Ring Q¹ is any        of the following (i) to (vii):        (i) a benzene ring optionally having one or two substituents        independently selected from the above Group A;        (ii) a pyridine ring optionally having one or two substituents        independently selected from the above Group A;        (iii) a 1,3-thiazole ring or a pyrazole ring (the 1,3-thiazole        ring or pyrazole ring optionally has one substituent        independently selected from the above Group A);        (iv) a cyclohexane ring optionally having one substituent        independently selected from the above Group A;        (v) a cyclohexene ring optionally having one substituent        independently selected from the above Group A;        (vi) a piperidine ring optionally having one substituent        independently selected from the above Group A; or        (vii) an indole ring optionally has one or two substituents        independently selected from the above Group B.        [17] The compound according to any one of [1] to [15], or a        pharmaceutically acceptable salt thereof, wherein m is 1, and        Ring Q¹ is any of the following formulas (12A) to (12H):

-   -   wherein    -   * is bonded to Z,    -   ** is bonded to the carbon atom to which R¹ is bonded,    -   R²⁷ is a hydrogen atom, a halogen atom, a C₁₋₆ alkoxy group,    -   or a C₁₋₆ alkyl group,    -   J is a nitrogen atom or CR²⁹,    -   R²⁹ is a halogen atom, and    -   R²⁸ is a hydrogen atom or a C₁₋₆ alkyl group.        [18] The compound according to any one of [1] to [15], or a        pharmaceutically acceptable salt thereof, wherein m is 1, and        Ring Q¹ is the following formula (13A) or (13B):

-   -   wherein    -   *is bonded to Z,    -   ** is bonded to the carbon atom to which R¹ is bonded, and    -   R³⁰ is a hydrogen atom, a fluorine atom, a methyl group, or a        methoxy group.        [19] The compound according to any one of [1] to [18], or a        pharmaceutically acceptable salt thereof, wherein Ring Q² is any        of the following (i) to (vii):        (i) a benzene ring optionally having one to three substituents        independently selected from the above Group C;        (ii) a pyridine ring optionally having one to three substituents        independently selected from the above Group C;        (iii) a pyridazine ring, a pyrazine ring or a pyrimidine ring        (the pyridazine ring, pyrazine ring or pyrimidine ring        optionally has one to three substituents independently selected        from the above Group C);        (iv) a pyrazole ring, an imidazole ring, a 1,3-thiazole ring, a        1,3-oxazole ring or a 4H-1,2,4-triazole ring (the pyrazole ring,        imidazole ring, 1,3-thiazole ring, 1,3-oxazole ring or        4H-1,2,4-triazole ring optionally has one substituent        independently selected from the above Group C);        (v) an isoquinoline ring, an indazole ring, a benzimidazole        ring, a 1H-pyrrolo[2,3-c]pyridine ring, a        1H-pyrrolo[3,2-c]pyridine ring, a furo[3,2-b]pyridine ring, a        1H-pyrazolo[3,4-c]pyridine ring or an indoline ring (the        isoquinoline ring, indazole ring, benzimidazole ring,        1H-pyrrolo[2,3-c]pyridine ring, 1H-pyrrolo[3,2-c]pyridine ring,        furo[3,2-b]pyridine ring, 1H-pyrazolo[3,4-c]pyridine ring or        indoline ring optionally has one or two substituents        independently selected from the above Group D);        (vi) a pyrrolidine ring, a piperidine ring, a morpholine ring or        an azepane ring (the pyrrolidine ring, piperidine ring,        morpholine ring or azepane ring optionally has one substituent        independently selected from the above Group E); or        (vii) a cyclohexane ring optionally having one substituent        independently selected from the above Group E.        [20] The compound according to any one of [1] to [18], or a        pharmaceutically acceptable salt thereof, wherein        W is the above formula (4A); and        Ring Q² is any of the following formulas (14A) to (14F):

-   -   wherein    -   * indicates a bonding site,    -   T is CH or a nitrogen atom,    -   R³¹ is a hydrogen atom, a C₁₋₆ alkoxy group, a halogeno C₁₋₆        alkoxy group, or a (²H3)methoxy group,    -   R³² is a hydrogen atom, a C₁₋₆ alkyl group, a halogen atom, a        C₁₋₆ alkoxy group, a cyano group, a di(C₁₋₆ alkyl)amino group, a        halogeno C₁₋₆ alkyl group, a C₁₋₆ alkylamino group, a C₁₋₆        alkylsulfonyl group, a C₁₋₆ alkoxy C₁₋₆ alkoxy group, a halogeno        C₁₋₆ alkoxy group, a hydroxy C₁₋₆ alkyl group, a C₁₋₆ alkyl        (2-C₃₋₆ alkenoyl)amino group, a (²H3)methoxy group, or    -   a bis[(²H3)methyl]amino group, or    -   R³¹ and R³² are taken together to form an ethylenedioxy group,    -   R³³ and R³⁵ are each independently a hydrogen atom, a halogen        atom, a C₁₋₆ alkoxy group, a C₁₋₆ alkyl(C₁₋₆ alkylsulfonyl)amino        group, a (C₁₋₆ alkyl)carbamoyl group, a di(C₁₋₆ alkyl)sulfamoyl        group, a 2-C₃₋₆ alkenoylamino group,    -   or a C₁₋₆ alkylsulfonyl C₁₋₆ alkyl group,    -   R³⁴ is a hydrogen atom or a halogen atom,    -   R³⁶ is a halogen atom,    -   R³⁷ is a C₁₋₆ alkoxy group,    -   R³⁸ is a halogen atom,    -   R³⁹ is a C₁₋₆ alkyl group or a C₁₋₆ alkylsulfonyl group,    -   R⁴⁰ is a C₁₋₆ alkyl group or a C₁₋₆ alkylsulfonyl group,    -   U¹ is CH or a nitrogen atom,    -   U² is CR⁴¹ or a nitrogen atom, and    -   R⁴¹ is a hydrogen atom or a halogen atom.        [21] The compound according to any one of [1] to [18], or a        pharmaceutically acceptable salt thereof, wherein W is the above        formula (4A); and        Ring Q² is any of the following formulas (15A) to (15C):

-   -   wherein    -   * indicates a bonding site,    -   R⁴² is a methyl group, a chlorine atom, a methoxy group, a cyano        group, a dimethylamino group, or a    -   bis [(²H3)methyl] amino group,    -   R⁴³ is a methoxy group or a (²H3)methoxy group, and    -   R⁴⁴ is a chlorine atom, a methoxy group, a methoxyethoxy group,        a dimethylamino group, a difluoromethoxy group, or a        (²H₃)methoxy group.        [22] The compound according to any one of [1] to [18], or a        pharmaceutically acceptable salt thereof, wherein        W is the above formula (4A); and        Ring Q² is any of the following formulas (16A) to (16G):

-   -   wherein * indicates a bonding site.        [23] The compound according to any one of [1] to [18], or a        pharmaceutically acceptable salt thereof, wherein        W is the above formula (4B); and        Ring Q² is the following formula (17A) or (17B):

-   -   wherein    -   * is bonded to Y, and    -   ** is bonded to Z.        [24] The compound according to any one of [1] to [19] and [23],        or a pharmaceutically acceptable salt thereof, wherein W is the        above formula (4B); and        Ring Q³ is any of the following formulas (18A) to (18D):

-   -   wherein    -   * indicates a bonding site,    -   R⁴⁵ is a hydrogen atom or a halogen atom,    -   R⁴⁶ is a C₁₋₆ alkylsulfonyl group, and    -   V is a nitrogen atom or CH.        [25] The compound according to any one of [1] to [19] and [23],        or a pharmaceutically acceptable salt thereof, wherein        W is the above formula (4B); and        Ring Q³ is a phenyl group, an azetidin-1-yl group, a 3-pyridyl        group, a 6-chloro-3-pyridyl group, a tetrahydropyran-3-yl group,        or a 1-methylsulfonyl-4-piperidyl group.        [26] The compound according to any one of [1] to [19] and [23]        to [25], or a pharmaceutically acceptable salt thereof, wherein        W is the above formula (4B); and        Y is a single bond or an oxygen atom.        [27] The compound according to any one of [1] to [26], or a        pharmaceutically acceptable salt thereof, wherein Z is a single        bond, —NH—, an oxygen atom, —SO₂—, —CH₂—, *—CH₂—NHC(═O)—**,        *—CH₂CH₂—O—**, or *—CH₂—NH—**, wherein * is bonded to Ring Q²,        and ** is bonded to Ring Q¹.        [28] The compound according to any one of [1] to [26], or a        pharmaceutically acceptable salt thereof, wherein Z is a single        bond.        [29] The compound according to [1], or a pharmaceutically        acceptable salt thereof, wherein        R¹ is a hydrogen atom;        R² is a hydrogen atom;        the moiety represented by the following formula (5) is any of        the following formulas (9A) to (9C):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded, and    -   ** is bonded to the nitrogen atom to which R⁵ is bonded;        R⁵ is a methyl group;        R⁶ is a hydrogen atom, a methyl group, a chlorine atom, a        methoxy group, an amino group, or a methylamino group;        R⁷ and R⁸ are taken together with the carbon atom to which R⁷ is        bonded and the carbon atom to which R⁸ is bonded to form the        following formula (10A):

-   -   wherein    -   the dotted circle indicates that the ring is aromatic,    -   the carbon atom marked with a is the carbon atom to which R⁸ is        bonded, and    -   the carbon atom marked with b is the carbon atom to which R⁷ is        bonded, or        R⁷ is a hydrogen atom, and R⁸ is the following formula (11A) or        (11B):

-   -   wherein    -   * indicates a bonding site,    -   R²⁵ is a diisopropylcarbamoyl group, a 4-isopropylpyrimidin-5-yl        group, a 2-isopropylphenyl group, or a 1-isopropylpyrazol-5-yl        group, and    -   R²⁶ is a diisopropylcarbamoyl group;        m is 1;        Ring Q¹ is the following formula (13A) or (13B):

-   -   wherein    -   * is bonded to Z,    -   ** is bonded to the carbon atom to which R¹ is bonded, and    -   R³⁰ is a hydrogen atom, a fluorine atom, a methyl group, or    -   a methoxy group;        W is the above formula (4A), and        Ring Q² is any of the following formulas (15A) to (15C):

-   -   wherein    -   * indicates a bonding site,    -   R⁴² is a methyl group, a chlorine atom, a methoxy group, a cyano        group, a dimethylamino group, or a    -   bis [(²H₃)methyl] amino group,    -   R⁴³ is a methoxy group or a (²H₃)methoxy group, and    -   R⁴⁴ is a chlorine atom, a methoxy group, a methoxyethoxy group,        a dimethylamino group, a difluoromethoxy group, or a        (²H₃)methoxy group, or        W is the above formula (4B),        Ring Q² is the following formula (17A) or (17B):

-   -   wherein    -   * is bonded to Y, and    -   ** is bonded to Z,        Ring Q³ is a phenyl group, an azetidin-1-yl group, a 3-pyridyl        group, a 6-chloro-3-pyridyl group, a tetrahydropyran-3-yl group,        or a 1-methylsulfonyl-4-piperidyl group, and        Y is a single bond or an oxygen atom; and        Z is a single bond.        [30] The compound according to [1], or a pharmaceutically        acceptable salt thereof, wherein        R¹ is a hydrogen atom;        R² is a hydrogen atom;        the moiety represented by the following formula (5) is any of        the following formulas (9A) to (9C):

-   -   wherein    -   * is bonded to the nitrogen atom to which R² is bonded, and    -   ** is bonded to the nitrogen atom to which R⁵ is bonded;        R⁵ is a methyl group;        R⁶ is a hydrogen atom, a methyl group, a chlorine atom, a        methoxy group, an amino group, or a methylamino group;        R⁷ and R⁸ are taken together with the carbon atom to which R⁷ is        bonded and the carbon atom to which R⁸ is bonded to form the        following formula (10A):

-   -   wherein    -   the dotted circle indicates that the ring is aromatic,    -   the carbon atom marked with a is the carbon atom to which R⁸ is        bonded, and    -   the carbon atom marked with b is the carbon atom to which R⁷ is        bonded;        m is 1;        Ring Q¹ is the following formula (13A) or (13B):

wherein

-   -   * is bonded to Z,    -   ** is bonded to the carbon atom to which R¹ is bonded, and    -   R³⁰ is a hydrogen atom, a fluorine atom, a methyl group, or    -   a methoxy group;        W is the above formula (4A); and        Ring Q² is any of the following formulas (16A) to (16G):

-   -   wherein * indicates a bonding site; and        Z is a single bond.        [31] Any compound selected from the following group, or a        pharmaceutically acceptable salt thereof:

-   5-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-3-methoxypyridine-2-carbonitrile,

-   (1R,2S,4R)-4-[({4-[1-(methanesulfonyl)-1H-indazol-4-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(5-methoxy-6-methylpyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(6-chloro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(6-fluoro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(6-chloro-5-methoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   2-[(4-{[(1S,2R,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-5-fluoro-N,N-di(propan-2-yl)benzamide,

-   (1R,2S,4R)-2-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)cyclopentan-1-ol,

-   (1R,3S)—N³-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-N′-methyl-N′-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine,

-   (1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   6-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-4-methoxypyridazine-3-carbonitrile,

-   (1S,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-[({4-[5-methoxy-6-(2-methoxyethoxy)pyridazin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(4,5-dimethoxypyridin-2-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-[({4-[6-(difluoromethoxy)-5-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-{[(4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}phenyl)methyl]amino}-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,

-   (1R,2S,4R)-4-({[4-(6-{bis[(²H₃)methyl]amino}-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,    and

-   (1R,2S,4R)-4-{[(4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}phenyl)methyl]amino}-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol.    [32] An inhibitor of the interaction between menin and one or more    proteins selected from the group consisting of MLL1, MLL2, a MLL    fusion protein and a MLL partial tandem duplication protein, which    comprises, as an active ingredient, the compound according to any    one of [1] to [31], or a pharmaceutically acceptable salt thereof.    [33] A pharmaceutical composition comprising the compound according    to any one of [1] to [31], or a pharmaceutically acceptable salt    thereof and a pharmaceutically acceptable carrier.    [34] The pharmaceutical composition according to [33], for the    treatment and/or prophylaxis of diseases that can be treated and/or    prevented by inhibiting the interaction between menin and one or    more proteins selected from the group consisting of MLL1, MLL2, a    MLL fusion protein and a MLL partial tandem duplication protein.    [35] The pharmaceutical composition according to [33], for the    treatment and/or prophylaxis of diabetes.    [36] The pharmaceutical composition according to [33], for the    treatment and/or prophylaxis of cancer.    [37] The pharmaceutical composition according to [36], wherein the    cancer is blood cancer, prostate cancer, breast cancer, hepatoma or    pediatric glioma.    [38] The pharmaceutical composition according to [36], wherein the    cancer is blood cancer.    [39] The pharmaceutical composition according to [38], wherein the    blood cancer is acute myelogenous leukemia (AML) or acute    lymphocytic leukemia (ALL).    [40] A method for treating and/or preventing diabetes, comprising    administering the compound according to any one of [1] to [31], or a    pharmaceutically acceptable salt thereof.    [41] A method for treating and/or preventing cancer, comprising    administering the compound according to any one of [1] to [31], or a    pharmaceutically acceptable salt thereof.    [42] The compound according to any one of [1] to [31], or a    pharmaceutically acceptable salt thereof, for use in the treatment    and/or prophylaxis of cancer.    [43] Use of the compound according to any one of [1] to [31], or a    pharmaceutically acceptable salt thereof, for the manufacture of a    medicament for the treatment and/or prophylaxis of cancer.    [44] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,    or a pharmaceutically acceptable salt thereof.    [45] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    succinate.    [46] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    benzenesulfonate.    [47] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    maleate.    [48] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    fumarate.    [49] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,    or a pharmaceutically acceptable salt thereof.    [50] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    fumarate.    [51] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    mucate.    [52] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    adipate.    [53] The compound according to [1], which is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    succinate.    [54] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    succinate, having at least five peaks at diffraction angles (2θ)    selected from 4.66±0.2, 7.02±0.2, 14.10±0.2, 16.68±0.2, 17.46±0.2,    18.68±0.2, 21.34±0.2, 24.52±0.2, 25.54±0.2 and 28.22±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [55] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    benzenesulfonate, having at least five peaks at diffraction angles    (2θ) selected from 10.92±0.2, 11.70±0.2, 12.40±0.2, 15.00±0.2,    17.38±0.2, 18.16±0.2, 22.18±0.2, 22.62±0.2, 23.86±0.2 and 24.20±0.2    in a powder X-ray diffraction diagram obtained through irradiation    with copper Kα line (λ=1.54 angstroms).    [56] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    maleate, having at least five peaks at diffraction angles (2θ)    selected from 4.64±0.2, 7.02±0.2, 7.46±0.2, 11.14±0.2, 14.04±0.2,    16.76±0.2, 18.54±0.2, 19.76±0.2, 21.26±0.2 and 22.62±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [57] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol    fumarate, having at least five peaks at diffraction angles (2θ)    selected from 4.80±0.2, 7.94±0.2, 9.66±0.2, 11.56±0.2, 14.56±0.2,    17.62±0.2, 18.14±0.2, 20.46±0.2, 21.36±0.2 and 24.46±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [58] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,    having at least five peaks at diffraction angles (2θ) selected from    7.14±0.2, 8.76±0.2, 12.26±0.2, 14.30±0.2, 17.52±0.2, 23.40±0.2,    24.40±0.2, 24.86±0.2, 25.34±0.2 and 25.90±0.2 in a powder X-ray    diffraction diagram obtained through irradiation with copper Kα line    (λ=1.54 angstroms).    [59] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    fumarate, having at least five peaks at diffraction angles (2θ)    selected from 8.06±0.2, 12.22±0.2, 12.52±0.2, 15.14±0.2, 17.54±0.2,    18.56±0.2, 20.08±0.2, 23.48±0.2, 24.28±0.2 and 25.00±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [60] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    mucate, having at least five peaks at diffraction angles (2θ)    selected from 6.56±0.2, 9.44±0.2, 9.94±0.2, 13.20±0.2, 18.22±0.2,    18.86±0.2, 19.60±0.2, 22.68±0.2, 25.10±0.2 and 28.70±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [61] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    adipate, having at least five peaks at diffraction angles (2θ)    selected from 5.88±0.2, 6.20±0.2, 9.18±0.2, 10.34±0.2, 12.50±0.2,    13.70±0.2, 15.66±0.2, 17.82±0.2, 18.48±0.2 and 22.16±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [62] A crystal of the compound according to [1], wherein the    compound is    (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol    succinate, having at least five peaks at diffraction angles (2θ)    selected from 4.60±0.2, 6.60±0.2, 7.74±0.2, 8.02±0.2, 9.26±0.2,    11.16±0.2, 12.00±0.2, 12.44±0.2, 13.22±0.2 and 19.66±0.2 in a powder    X-ray diffraction diagram obtained through irradiation with copper    Kα line (λ=1.54 angstroms).    [63] An inhibitor of the interaction between menin and one or more    proteins selected from the group consisting of MLL1, MLL2, a MLL    fusion protein and a MLL partial tandem duplication protein, which    comprises, as an active ingredient, the compound according to any    one of [1] to [31] and [44] to [53], or a pharmaceutically    acceptable salt thereof, or the crystal according to any one of [54]    to [62].    [64] A pharmaceutical composition comprising the compound according    to any one of [1] to [31] and [44] to [53], or a pharmaceutically    acceptable salt thereof, or the crystal according to any one of [54]    to [62], and a pharmaceutically acceptable carrier.    [65] The pharmaceutical composition according to [64], for the    treatment and/or prophylaxis of diseases that can be treated and/or    prevented by inhibiting the interaction between menin and one or    more proteins selected from the group consisting of MLL1, MLL2, a    MLL fusion protein and a MLL partial tandem duplication protein.    [66] The pharmaceutical composition according to [64], for the    treatment and/or prophylaxis of diabetes.    [67] The pharmaceutical composition according to [64], for the    treatment of cancer.    [68] The pharmaceutical composition according to [67], wherein the    cancer is blood cancer, prostate cancer, breast cancer, hepatoma or    pediatric glioma.    [69] The pharmaceutical composition according to [67], wherein the    cancer is blood cancer.    [70] The pharmaceutical composition according to [69], wherein the    blood cancer is acute myelogenous leukemia (AML) or acute    lymphocytic leukemia (ALL).    [71] A method for treating and/or preventing diabetes, comprising    administering the compound according to any one of [1] to [31] and    [44] to [53], or a pharmaceutically acceptable salt thereof, or the    crystal according to any one of [54] to [62].    [72] A method for treating cancer, comprising administering the    compound according to any one of [1] to [31] and [44] to [53], or a    pharmaceutically acceptable salt thereof, or the crystal according    to any one of [54] to [62].    [73] The compound according to any one of [1] to [31] and [44] to    [53], or a pharmaceutically acceptable salt thereof, or the crystal    according to any one of [54] to [62], for use in the treatment of    cancer.    [74] Use of the compound according to any one of [1] to [31] and    [44] to [53], or a pharmaceutically acceptable salt thereof, or the    crystal according to any one of [54] to [62], for the manufacture of    a medicament for the treatment of cancer.    [75] The pharmaceutical composition according to [69], wherein the    blood cancer is acute myelogenous leukemia (AML) with NPM1 mutation.    [76] A pharmaceutical composition comprising one drug selected from    the group consisting of a Bcl-2 inhibitor, a DNA methyltransferase    inhibitor and a pyrimidine antimetabolite, and the compound    according to any one of [1] to [31] and [44] to [53], or a    pharmaceutically acceptable salt thereof, or the crystal according    to any one of [54] to [62], which are administered in combination.    [77] The pharmaceutical composition according to [76], wherein the    one drug selected from the group consisting of a Bcl-2 inhibitor, a    DNA methyltransferase inhibitor and a pyrimidine antimetabolite, and    the compound according to any one of [1] to [31] and [44] to [53],    or a pharmaceutically acceptable salt thereof, or the crystal    according to any one of [54] to [62] are separately comprised as    active ingredients in different formulations and administered at the    same time or different times.    [78] The pharmaceutical composition according to [76], wherein the    one drug selected from the group consisting of a Bcl-2 inhibitor, a    DNA methyltransferase inhibitor and a pyrimidine antimetabolite, and    the compound according to any one of [1] to [31] and [44] to [53],    or a pharmaceutically acceptable salt thereof, or the crystal    according to any one of [54] to [62] are comprised in a single    formulation.    [79] The pharmaceutical composition according to any one of [76] to    [78], wherein the drug selected from the group consisting of a Bcl-2    inhibitor, a DNA methyltransferase inhibitor and a pyrimidine    antimetabolite is Venetoclax.    [80] The pharmaceutical composition according to any one of [76] to    [78], wherein the one drug selected from the group consisting of a    Bcl-2 inhibitor, a DNA methyltransferase inhibitor and a pyrimidine    antimetabolite is Azacitidine.    [81] The pharmaceutical composition according to any one of [76] to    [78], wherein the one drug selected from the group consisting of a    Bcl-2 inhibitor, a DNA methyltransferase inhibitor and a pyrimidine    antimetabolite is Cytarabine.    [82] The method according to [72], wherein the compound according to    any one of [1] to [31] and [44] to [53], or a pharmaceutically    acceptable salt thereof, or the crystal according to any one of [54]    to [62] is administered in combination with one drug selected from    the group consisting of a Bcl-2 inhibitor, a DNA methyltransferase    inhibitor and a pyrimidine antimetabolite.    [83] The method according to [82], wherein the one drug selected    from the group consisting of a Bcl-2 inhibitor, a DNA    methyltransferase inhibitor and a pyrimidine antimetabolite, and the    compound according to any one of [1] to [31] and [44] to [53], or a    pharmaceutically acceptable salt thereof, or the crystal according    to any one of [54] to [62] are separately comprised as active    ingredients in different formulations and administered at the same    time or different times.    [84] The method according to [82], wherein the one drug selected    from the group consisting of a Bcl-2 inhibitor, a DNA    methyltransferase inhibitor and a pyrimidine antimetabolite, and the    compound according to any one of [1] to [31] and [44] to [53], or a    pharmaceutically acceptable salt thereof, or the crystal according    to any one of [54] to [62] are comprised in a single formulation.    [85] The compound according to any one of [1] to [31] and [44] to    [53], or a pharmaceutically acceptable salt thereof, or the crystal    according to any one of [54] to [62], which is administered in    combination with one drug selected from the group consisting of a    Bcl-2 inhibitor, a DNA methyltransferase inhibitor and a pyrimidine    antimetabolite.    [86] The compound or pharmaceutically acceptable salt thereof or    crystal according to [85], wherein the one drug selected from the    group consisting of a Bcl-2 inhibitor, a DNA methyltransferase    inhibitor and a pyrimidine antimetabolite, and the compound    according to any one of [1] to [31] and [44] to [53], or a    pharmaceutically acceptable salt thereof, or the crystal according    to any one of [54] to [62] are separately comprised as active    ingredients in different formulations and administered at the same    time or different times.    [87] The compound or pharmaceutically acceptable salt thereof or    crystal according to [85], wherein the one drug selected from the    group consisting of a Bcl-2 inhibitor, a DNA methyltransferase    inhibitor and a pyrimidine antimetabolite, and the compound    according to any one of [1] to [31] and [44] to [53], or a    pharmaceutically acceptable salt thereof, or the crystal according    to any one of [54] to [62] are comprised in a single formulation.    [88] The compound or pharmaceutically acceptable salt thereof or    crystal according to any one of [85] to [87], wherein the one drug    selected from the group consisting of a Bcl-2 inhibitor, a DNA    methyltransferase inhibitor and a pyrimidine antimetabolite is    Venetoclax.    [89] The compound or pharmaceutically acceptable salt thereof or    crystal according to any one of [85] to [87], wherein the one drug    selected from the group consisting of a Bcl-2 inhibitor, a DNA    methyltransferase inhibitor and a pyrimidine antimetabolite is    Azacitidine.    [90] The compound or pharmaceutically acceptable salt thereof or    crystal according to any one of [85] to [87], wherein the one drug    selected from the group consisting of a Bcl-2 inhibitor, a DNA    methyltransferase inhibitor and a pyrimidine antimetabolite is    Cytarabine.    [91] A composition for inducing differentiation of leukemia cells,    comprising the compound according to any one of [1] to [31] and [44]    to [53], or a pharmaceutically acceptable salt thereof, or the    crystal according to any one of [54] to [62].    [92] A method for inducing differentiation of leukemia cells,    comprising administering the compound according to any one of [1] to    [31] and [44] to [53], or a pharmaceutically acceptable salt    thereof, or the crystal according to any one of [54] to [62].

Effect of the Invention

The compound of the present invention or a pharmaceutically acceptablesalt thereof exhibits an inhibitory action on the interaction betweenmenin and an MLL protein. Specifically, administration of thepharmaceutical composition comprising the compound of the presentinvention or a pharmaceutically acceptable salt thereof and apharmaceutically acceptable carrier to a mammal (human, bovine, horse,swine, etc.) or a bird (chicken, etc.) can be employed for the treatmentand/or prophylaxis of diseases dependent on the interaction betweenmenin and an MLL protein. Examples of the disease dependent on theinteraction between menin and an MLL protein include cancers anddiabetes. Examples of the cancer include blood cancer, myelodysplasticsyndrome, prostate cancer, breast cancer, hepatoma and pediatric glioma,preferred is blood cancer, and more preferred are acute myelogenousleukemia (AML) and acute lymphocytic leukemia (ALL).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a powder X-ray diffraction diagram of the crystal obtained inExample 131. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 2 is a powder X-ray diffraction diagram of the crystal obtained inExample 132. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 3 is a powder X-ray diffraction diagram of the crystal obtained inExample 133. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 4 is a powder X-ray diffraction diagram of the crystal obtained inExample 134. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 5 is a powder X-ray diffraction diagram of the crystal obtained inExample 135. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 6 is a powder X-ray diffraction diagram of the crystal obtained inExample 136. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 7 is a powder X-ray diffraction diagram of the crystal obtained inExample 137. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 8 is a powder X-ray diffraction diagram of the crystal obtained inExample 138. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 9 is a powder X-ray diffraction diagram of the crystal obtained inExample 139. The vertical axis indicates the diffraction intensity(Intensity) in count/sec (cps) unit, and the horizontal axis indicatesthe value of the diffraction angle 2θ.

FIG. 10 is a graph showing the rate of the myeloid cell differentiationantigen Gr-1-expressing cells in living cells after treatment with thecompound of Example 25, 27, 26 or 22 for 7 days. The vertical axisindicates the percentage of the myeloid cell differentiation antigenGr-1-expressing cells in living cells, and the horizontal axis indicateseach compound and concentration (nM) thereof.

FIG. 11 is a graph showing the rate of the cKit-expressing cells inliving cells after treatment with the compound of Example 25, 27, 26 or22 for 7 days. The vertical axis indicates the percentage of thecKit-expressing cells in living cells, and the horizontal axis indicateseach compound and concentration (nM) thereof.

FIG. 12 is a graph showing the effects of the combined effect of thecompound of Example 25 and 5Aza on the in-vitro growth of human AML cellline MOLM-13 cells. The vertical axis indicates the cell growth (%), andthe horizontal axis indicates the concentration (nM) of the compound ofExample 25. The symbol black circle indicates the compound of Example 25alone, the symbol black triangle indicates the compound of Example25+5Aza (2.5 μM), the symbol black square indicates the compound ofExample 25+5Aza (5 μM), and the symbol x indicates the compound ofExample 25+5Aza (10 μM). The error bar indicates SD.

FIG. 13 is a graph showing the effects of the combined effect of thecompound of Example 25 and AraC on the in-vitro growth of human AML cellline MOLM-13 cells. The vertical axis indicates the cell growth (%), andthe horizontal axis indicates the concentration (nM) of the compound ofExample 25. The symbol black circle indicates the compound of Example 25alone, the symbol black triangle indicates the compound of Example25+AraC (25 nM), the symbol black square indicates the compound ofExample 25+AraC (50 nM), and the symbol x indicates the compound ofExample 25+AraC (100 nM). The error bar indicates SD.

FIG. 14 is a graph showing the effects of the combined effect of thecompound of Example 25 and Venetoclax on the in-vitro growth of humanAML cell line MOLM-13 cells. The vertical axis indicates the cellgrowth, and the horizontal axis indicates the concentration (nM) of thecompound of Example 25. The symbol black circle indicates the compoundof Example 25 alone, the symbol black triangle indicates the compound ofExample 25+Venetoclax (39 nM), the symbol black square indicates thecompound of Example 25+Venetoclax (78 nM), and the symbol x indicatesthe compound of Example 25+Venetoclax (156 nM). The error bar indicatesSD.

DESCRIPTION OF EMBODIMENTS

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which this invention belongs.

In the present invention, the “halogen atom” refers to a fluorine atom,a chlorine atom, a bromine atom or an iodine atom.

In the present invention, the “C₁₋₆ alkyl group” refers to a linear orbranched alkyl group having 1 to 6 carbon atoms. Examples thereofinclude a methyl group, an ethyl group, a n-propyl group, an isopropylgroup, a n-butyl group, a sec-butyl group, a tert-butyl group, an-pentyl group, an isopentyl group, a 2-methylbutyl group, a neo-pentylgroup, a 1-ethylpropyl group, a n-hexyl group, a 4-methylpentyl group, a3-methylpentyl group, a 2-methylpentyl group, a 1-methylpentyl group, a3,3-dimethylbutyl group, a 2,2-dimethylbutyl group, a 1,1-dimethylbutylgroup, a 1,2-dimethylbutyl group, a 1,3-dimethylbutyl group, a2,3-dimethylbutyl group, a 2-ethylbutyl group and the like.

In the present invention, the “C₁₋₆ alkoxy group” refers to a group inwhich the above “C₁₋₆ alkyl group” is bonded to an oxygen atom. Examplesthereof include a methoxy group, an ethoxy group, a n-propoxy group, anisopropoxy group, a n-butoxy group, a sec-butoxy group, a tert-butoxygroup, a n-pentoxy group, an isopentoxy group, a 2-methylbutoxy group, an-hexyloxy group and the like.

In the present invention, the (C₁₋₆ alkyl)carbamoyl group refers to agroup in which one hydrogen atom of a carbamoyl group is substitutedwith the above “C₁₋₆ alkyl group”. Examples thereof include amethylcarbamoyl group, an ethylcarbamoyl group, a propylcarbamoyl group,an isopropylcarbamoyl group, a sec-butylcarbamoyl group, a1-ethylpropylcarbamoyl group and the like.

In the present invention, the “di(C₁₋₆ alkyl)carbamoyl group” refers toa group in which two hydrogen atoms of a carbamoyl group are substitutedwith the same or different two of the above “C₁₋₆ alkyl groups”.Examples thereof include a dimethylcarbamoyl group, anethyl(methyl)carbamoyl group, a methyl(propyl)carbamoyl group, adiethylcarbamoyl group, a dipropylcarbamoyl group, adiisopropylcarbamoyl group, a sec-butyl(pentyl)carbamoyl group and thelike.

In the present invention, the “hydroxy C₁₋₆ alkyl group” refers to agroup in which one hydrogen atom of the above “C₁₋₆ alkyl group” issubstituted with a hydroxy group. Examples thereof include ahydroxymethyl group, a 1-hydroxyethyl group, a 2-hydroxyethyl group, a1-hydroxypropyl group, a 2-hydroxypropyl group, a 1-hydroxyisopropylgroup, a 1-hydroxybutyl group, a 2-hydroxybutyl group, a 1-hydroxypentylgroup, a 2-hydroxypentyl group, a 1-hydroxyhexyl group and the like.

In the present invention, the “hydroxy C₁₋₆ alkoxy group” refers to agroup in which one hydrogen atom of the above “C₁₋₆ alkoxy group” issubstituted with a hydroxy group. Examples thereof include ahydroxymethoxy group, a 2-hydroxyethoxy group, a 2-hydroxypropoxy group,a 3-hydroxypropoxy group, a 2-hydroxy-1-methyl-ethoxy group, a3-hydroxybutoxy group, a 2-hydroxybutoxy group, a 2-hydroxypentoxygroup, a 5-hydroxypentoxy group, a 4-hydroxyhexoxy group and the like.

In the present invention, the “C₁₋₆ alkylamino group” refers to a groupin which one hydrogen atom of an amino group is substituted with theabove “C₁₋₆ alkyl group”. Examples thereof include a methylamino group,an ethylamino group, a n-propylamino group, an isopropylamino group, an-butylamino group, a sec-butylamino group, a tert-butylamino group, an-pentylamino group and the like.

In the present invention, the “di(C₁₋₆ alkyl)amino group” refers to agroup in which two hydrogen atoms of an amino group are substituted withthe same or different two of the above “C₁₋₆ alkyl groups”. Examplesthereof include a dimethylamino group, a methyl(ethyl)amino group, amethyl(propyl)amino group [e.g., a N-methyl-N-(1-propyl)amino groupetc.], a methyl(butyl)amino group [e.g., a N-(1-butyl)-N-methylaminogroup etc.], a methyl(pentyl)amino group, a methyl(hexyl)amino group, adiethylamino group, an ethyl(propyl)amino group [e.g., aN-ethyl-N-(1-propyl)amino group etc.], an ethyl(butyl)amino group, adipropylamino group, a propyl(butyl)amino group, a dibutylamino group, adipentylamino group, a dihexylamino group and the like.

In the present invention, the “halogeno C₁₋₆ alkyl group” refers to agroup in which one to three hydrogen atoms of the above “C₁₋₆ alkylgroup” are substituted with the above “halogen atoms”. Examples thereofinclude a fluoromethyl group, a difluoromethyl group, a trifluoromethylgroup, a chloromethyl group, a dichloromethyl group, a trichloromethylgroup, a 1-fluoroethyl group, a 1-chloroethyl group, a 2-fluoroethylgroup, a 1,2-difluoropropyl group, a 2,2,2-trifluoroethyl group and thelike.

In the present invention, the “C₃₋₈ cycloalkyl group” refers to a 3- to8-membered monocyclic saturated hydrocarbon group (ring). Examplesthereof include a cyclopropyl group, a cyclobutyl group, a cyclopentylgroup, a cyclohexyl group, a cycloheptyl group and a cyclooctyl group.

In the present invention, the “C₃₋₈ cycloalkyl C₁₋₆ alkyl group” refersto a group in which one hydrogen atom of the above “C₁₋₆ alkyl group” issubstituted with the above “C₃₋₈ cycloalkyl group”. Examples thereofinclude a cyclopropylmethyl group, a 2-cyclobutylethyl group, a3-cyclopentylbutyl group, a 3-cycloheptyl-2-methyl-butyl group and thelike.

In the present invention, the “C₁₋₆ alkoxy C₁₋₆ alkyl group” refers to agroup in which one hydrogen atom of the above “C₁₋₆ alkyl group” issubstituted with the above “C₁₋₆ alkoxy group”. Examples thereof includea methoxymethyl group, an ethoxymethyl group, a n-propoxymethyl group,an isopropoxymethyl group, a 2-methoxyethyl group, a 2-ethoxyethylgroup, a 1-propoxyethyl group, a 1-isopropoxyethyl group and the like.

In the present invention, the “oxetanyl group” refers to an oxetan-3-ylgroup or an oxetan-2-yl group.

In the present invention, the “(C₁₋₆ alkyl)pyrimidinyl group” refers toa group in which one hydrogen atom of a pyrimidinyl group is substitutedwith the above “C₁₋₆ alkyl group”. Examples thereof include a4-isopropylpyrimidin-5-yl group, a 5-methylpyrimidin-2-yl group, a5-sec-butylpyrimidin-4-yl group, a 4-pentylpyrimidin-5-yl group and thelike.

In the present invention, the “(C₁₋₆ alkyl)phenyl group” refers to agroup in which one hydrogen atom of a phenyl group is substituted withthe above “C₁₋₆ alkyl group”. Examples thereof include a 3-tolyl group,a 2-ethylphenyl group, a 2-isopropylphenyl group, a4-(2,3-dimethylbutyl)phenyl group and the like.

In the present invention, the “(C₁₋₆ alkyl)pyrazolyl group” refers to agroup in which one hydrogen atom of a pyrazolyl group is substitutedwith the above “C₁₋₆ alkyl group”. Examples thereof include a3-methyl-1H-pyrazol-4-yl group, a 2-isopropylpyrazol-3-yl group, a4-isopentyl-1H-pyrazol-5-yl group, a 3-hexylpyrazol-1-yl group and thelike.

In the present invention, the “C₁₋₆ alkylsulfonyl group” refers to agroup in which the above “C₁₋₆ alkyl group” is bonded to a sulfur atomof a sulfonyl group. Examples thereof include a methylsulfonyl group, anethylsulfonyl group, a n-propylsulfonyl group, an isopropylsulfonylgroup, a n-butylsulfonyl group, a sec-butylsulfonyl group, atert-butylsulfonyl group, a n-pentylsulfonyl group and the like.

In the present invention, the “C₁₋₆ alkylene group” refers to a linearor branched alkylene group having 1 to 6 carbon atoms. Examples thereofinclude a methylene group, an ethylene group [—(CH₂)₂—], a trimethylenegroup [—(CH₂)₃—], a tetramethylene group, a pentamethylene group, ahexamethylene group, a methylmethylene group [—CH(CH₃)—], amethylethylene group [—CH(CH₃)CH₂— or —CH₂CH(CH₃)—], an ethylethylenegroup [—CH(CH₂CH₃)CH₂— or —CH₂CH(CH₂CH₃)—], a 1,2-dimethylethylene group[—CH(CH₃)CH(CH₃)—], a 1,1,2,2-tetramethylethylene group[—C(CH₃)₂C(CH₃)₂—] and the like.

In the present invention, the “vinylsulfonylamino(C₁₋₆ alkyl)carbamoylgroup” refers to a group in which one hydrogen atom of a carbamoyl groupis substituted with the following “vinylsulfonylamino(C₁₋₆ alkyl)group”. Examples thereof include a (vinylsulfonylamino)methylcarbamoylgroup, a 2-(vinylsulfonylamino)ethylcarbamoyl group, a3-(vinylsulfonylamino)propylcarbamoyl group, a2-[(vinylsulfonylamino)methyl]butylcarbamoyl group and the like.

In the present invention, the “vinylsulfonylamino(C₁₋₆ alkyl) group”refers to a group in which one hydrogen atom of the above “C₁₋₆ alkylgroup” is substituted with the following “vinylsulfonylamino group”.Examples thereof include a (vinylsulfonylamino)methyl group, a1-(vinylsulfonylamino)ethyl group, a 3-(vinylsulfonylamino)propyl group,a 3-methyl-4-(vinylsulfonylamino)butyl group and the like.

In the present invention, the “vinylsulfonylamino group” refers to agroup in which one hydrogen atom of an amino group is substituted withthe following “vinylsulfonyl group”.

In the present invention, the “vinylsulfonyl group” refers to a group inwhich a vinyl group is bonded to a sulfur atom of a sulfonyl group.

In the present invention, the “prop-2-enoylamino(C₁₋₆ alkyl)carbamoylgroup” refers to one hydrogen atom of a carbamoyl group is substitutedwith the following “prop-2-enoylamino(C₁₋₆ alkyl) group”. Examplesthereof include a (prop-2-enoylamino)methylcarbamoyl group, a2-(prop-2-enoylamino)ethylcarbamoyl group, a3-(prop-2-enoylamino)propylcarbamoyl group, a[2-methyl-3-(prop-2-enoylamino)propyl]carbamoyl group, a2-(prop-2-enoylamino)pentylcarbamoyl group and the like.

In the present invention, the “prop-2-enoylamino(C₁₋₆ alkyl) group”refers to a group in which one hydrogen atom of the above “C₁₋₆ alkylgroup” is substituted with the following “prop-2-enoylamino group”.Examples thereof include a (prop-2-enoylamino)methyl group, a2-(prop-2-enoylamino)ethyl group, a 3-(prop-2-enoylamino)propyl group, a2-methyl-3-(prop-2-enoylamino)propyl group, a 4-(prop-2-enoylamino)butylgroup, a 6-(prop-2-enoylamino)hexyl group and the like.

In the present invention, the “prop-2-enoylamino group” refers to agroup in which one hydrogen atom of an amino group is substituted withthe following “prop-2-enoyl group”.

In the present invention, the “prop-2-enoyl group” refers to a group inwhich a vinyl group is bonded to a carbon atom of a carbonyl group.

In the present invention, the “C₁₋₆ alkyl (C₁₋₆ alkylsulfonyl)aminogroup” refers to a group in which two hydrogen atoms of an amino groupare substituted with the above “C₁₋₆ alkyl group” and the above “C₁₋₆alkylsulfonyl”. Examples thereof include a methyl(methylsulfonyl)aminogroup, an ethyl(isopropylsulfonyl)amino group, abutylsulfonyl(propyl)amino group, a hexylsulfonyl(isobutyl)amino groupand the like.

In the present invention, the “C₁₋₆ alkoxy C₁₋₆ alkoxy group” refers toa group in which one hydrogen atom of the above “C₁₋₆ alkoxy group” issubstituted with the above “C₁₋₆ alkoxy group”. Examples thereof includea methoxymethoxy group, an ethoxymethoxy group, a n-propoxymethoxygroup, an isopropoxymethoxy group, a methoxyethoxy group, anethoxyethoxy group, a n-propoxyethoxy group, an isopropoxyethoxy groupand the like.

In the present invention, the “halogeno C₁₋₆ alkoxy group” refers to agroup in which one or two hydrogen atoms of the above “C₁₋₆ alkoxygroup” are substituted with the above “halogen atoms”. Examples thereofinclude a fluoromethoxy group, a difluoromethoxy group, a chloromethoxygroup, a dichloromethoxy group, a 1-fluoroethoxy group, a 1-chloroethoxygroup, a 2-fluoroethoxy group, a 1,2-difluoropropoxy group and the like.

In the present invention, the “C₁₋₆ alkylsulfonyl C₁₋₆ alkyl group”refers to a group in which one hydrogen atom of the above “C₁₋₆ alkylgroup” is substituted with the above “C₁₋₆ alkylsulfonyl group”.Examples thereof include a methylsulfonylmethyl group, amethylsulfonylethyl group, an ethylsulfonylmethyl group, an-propylsulfonylmethyl group, an isopropylsulfonylmethyl group, an-butylsulfonylmethyl group, a sec-butylsulfonylmethyl group, atert-butylsulfonylmethyl group, a tert-butylsulfonylethyl group, an-pentylsulfonylmethyl group and the like.

In the present invention, the “di(C₁₋₆ alkyl)sulfamoyl group” refers toa group in which two hydrogen atoms of the following “sulfamoyl group”are substituted with the same or different two of the above “C₁₋₆ alkylgroups”. Examples thereof include a dimethylsulfamoyl group, anethyl(methyl)sulfamoyl group, an ethyl(isopropyl)sulfamoyl group, adibutylsulfamoyl group, a hexyl(isopentyl)sulfamoyl group and the like.

In the present invention, the “sulfamoyl group” refers to an amino groupis bonded to a sulfur atom of a sulfonyl group.

In the present invention, the “C₁₋₆ alkylenedioxy group” refers to agroup in which two hydrogen atoms of the above “C₁₋₆ alkyl group” aresubstituted with oxy groups (—O—). Examples thereof include amethylenedioxy group (—OCH₂O—), an ethylenedioxy group [—O(CH₂)₂O-], atrimethylenedioxy group [—O(CH₂)₃O—], a methylethylenedioxy group[—OCH(CH₃)CH₂O— or —OCH₂CH(CH₃)O—] and the like.

In the present invention, the “2-C₃₋₆ alkenoylamino group” refers to agroup in which one hydrogen atom of an amino group is substituted withthe following “2-C₃₋₆ alkenoyl group”. Examples thereof include aprop-2-enoylamino group, a 2-methylprop-2-enoylamino group, a3-methylbuta-2-enoylamino group, an [(E)-penta-2-enoyl]amino group, an[(E)-3-methylpenta-2-enoyl]amino group and the like.

In the present invention, the “2-C₃₋₆ alkenoyl group” refers to a groupin which the following “1-C₂₋₅ alkenyl group” is bonded to a carbon atomof a carbonyl group. Examples thereof include a prop-2-enoyl group, an(E)-buta-2-enoyl group, a 3-methylbuta-2-enoyl group, an(E)-hexa-2-enoyl group, a 2-methylprop-2-enoyl group, a3-methyl-2-methylene-butanoyl group and the like.

In the present invention, the “1-C₂₋₅ alkenyl group” refers to a linearor branched alkenyl group having 2 to 5 carbon atoms (the bonding siteof the alkenyl group is present on the unsaturated carbon atom).Examples thereof include an (E)-prop-1-enyl group, a 2-methylprop-1-enylgroup, an (E)-pent-1-enyl group, an isopropenyl group, a1-methylenebutyl group, a (Z)-1-ethylprop-1-enyl group and the like.

In the present invention, the “C₁₋₆ alkyl (2-C₃₋₆ alkenoyl)amino group”refers to a group in which two hydrogen atoms of an amino group aresubstituted with the above “C₁₋₆ alkyl group” and the above “2-C₃₋₆alkenoyl group”. Examples thereof include a methyl(prop-2-enoyl)aminogroup, an ethyl(2-methylprop-2-enoyl)amino group, a3-methylbuta-2-enoyl(propyl)amino group, anisopropyl-[(E)-penta-2-enoyl]amino group and the like.

In the present invention, the “(²H₃)methoxy group” refers to a group inwhich three hydrogen atoms of a methoxy group are all substituted withdeuteriums (²H; D).

In the present invention, the “bis[(²H₃)methyl]amino group” refers to agroup in which six hydrogen atoms of a dimethylamino group are allsubstituted with deuteriums (²H; D).

In the present invention, the “when a ring has a oxo group” refers to acase where an oxo group is bonded to a ring-constituting atom. Forexample, when a pyridine ring has an oxo group, examples thereof includea 1H-pyridin-2-one ring, a 4H-pyridin-3-one and the like, and when apyridazine ring has an oxo group, examples thereof include a1H-pyridazin-6-one ring and the like.

In the present invention, the “6-membered aromatic ring optionallycontaining one nitrogen atom in the ring” refers to a 6-memberedmonocyclic aromatic ring optionally containing one nitrogen atom as aring-constituting atom, besides a carbon atom. Examples thereof includea benzene ring and a pyridine ring. The “6-membered aromatic ringoptionally containing one nitrogen atom in the ring” for Ring Q¹ ispreferably a benzene ring or a pyridine ring, more preferably a benzenering. The “6-membered aromatic ring optionally containing one nitrogenatom in the ring” for Ring Q² is preferably a benzene ring or a pyridinering, more preferably a pyridine ring. The “6-membered aromatic ringoptionally containing one nitrogen atom in the ring” for Ring Q³ ispreferably a benzene ring or a pyridine ring, more preferably a pyridinering.

In the present invention, the “5-membered aromatic heterocyclecontaining, in the ring, one or two heteroatoms independently selectedfrom the group consisting of a nitrogen atom and a sulfur atom” refersto a 5-membered monocyclic aromatic ring containing one or twoheteroatoms (a nitrogen atom or a sulfur atom) as a ring-constitutingatom, besides a carbon atom. Examples thereof include a thiophene ring,a 1,2-thiazole ring, a 1,3-thiazole ring, a pyrrole ring, a pyrazolering and an imidazole ring. The “5-membered aromatic heterocyclecontaining, in the ring, one or two heteroatoms independently selectedfrom the group consisting of a nitrogen atom and a sulfur atom” for RingQ¹ is preferably a 1,3-thiazole ring or a pyrazole ring.

In the present invention, the “C₃₋₈ cycloalkane ring” refers to a 3- to8-membered monocyclic saturated hydrocarbon ring. Examples thereofinclude a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, acyclohexane ring, a cycloheptane ring and a cyclooctane ring. The “C₃₋₈cycloalkane ring” for Ring Q¹ is preferably a cyclohexane ring. The“C₃₋₈ cycloalkane ring” for Ring Q² is preferably a cyclohexane ring.

In the present invention, the “C₄₋₈ cycloalkene ring” refers to a 4- to8-membered monocyclic unsaturated hydrocarbon ring having one doublebond in the ring. Examples thereof include a cyclobutene ring, acyclopentene ring, a cyclohexene ring, a cycloheptene ring and acyclooctene ring. The “C₄₋₈ cycloalkene ring” for Ring Q¹ is preferablya cyclohexene ring.

In the present invention, the “4- to 8-membered saturated heterocyclecontaining one nitrogen atom in the ring” refers to a 4- to 8-memberedmonocyclic saturated ring containing one nitrogen atom as aring-constituting atom, besides a carbon atom. Examples thereof includean azetidine ring, a pyrrolidine ring, a piperidine ring, an azepanering and an azocane ring. The “4- to 8-membered saturated heterocyclecontaining one nitrogen atom in the ring” for Ring Q¹ is preferably apiperidine ring.

In the present invention, the “9-membered bicyclic aromatic heterocyclecontaining one nitrogen atom in the ring” refers to a 9-memberedbicyclic fused aromatic ring containing one nitrogen atom as aring-constituting atom, besides a carbon atom. Examples thereof includean indole ring, an isoindole ring, an indolizine ring and the like. The“9-membered bicyclic aromatic heterocycle containing one nitrogen atomin the ring” for Ring Q¹ is preferably an indole ring.

In the present invention, the “6-membered aromatic heterocyclecontaining two nitrogen atoms in the ring” refers to a 6-memberedmonocyclic aromatic ring containing two nitrogen atoms as aring-constituting atom, besides a carbon atom. Examples thereof includea pyridazine ring, a pyrazine ring and a pyrimidine ring. The“6-membered aromatic heterocycle containing two nitrogen atoms in thering” for Ring Q² is preferably a pyridazine ring, a pyrazine ring or apyrimidine ring.

In the present invention, the “5-membered aromatic heterocyclecontaining, in the ring, one to three heteroatoms independently selectedfrom the group consisting of a nitrogen atom, an oxygen atom and asulfur atom” refers to a 5-membered monocyclic aromatic ring containingone to three heteroatoms (a nitrogen atom, an oxygen atom or a sulfuratom) as a ring-constituting atom, besides a carbon atom. Examplesthereof include a pyrrole ring, a furan ring, a thiophene ring, animidazole ring, a pyrazole ring, a 1,2-oxazole ring, a 1,3-oxazole ring,a 1,2-thiazole ring, a 1,3-thiazole ring, a 4H-1,2,4-triazole ring, a1H-1,2,3-triazole ring, a 1,2,4-oxadiazole ring, a 1,3,4-thiadiazolering and the like. The “5-membered aromatic heterocycle containing, inthe ring, one to three heteroatoms independently selected from the groupconsisting of a nitrogen atom, an oxygen atom and a sulfur atom” forRing Q² is preferably an imidazole ring, a pyrazole ring, a 1,3-thiazolering, a 1,3-oxazole ring or a 4H-1,2,4-triazole ring.

In the present invention, the “9- or 10-membered bicyclic aromatic orpartially unsaturated heterocycle containing, in the ring, one to threeheteroatoms independently selected from the group consisting of anitrogen atom and an oxygen atom” refers to a ring derived from a 9- or10-membered bicyclic fused aromatic ring containing one to threeheteroatoms (a nitrogen atom or an oxygen atom) as a ring-constitutingatom, besides a carbon atom, which optionally having a saturated bond ina part of the bicyclic ring. Examples thereof include an indole ring, abenzofuran ring, an indazole ring, a benzimidazole ring, a1H-pyrrolo[2,3-c]pyridine ring, a 1H-pyrrolo[3,2-c]pyridine ring, afuro[3,2-b]pyridine ring, a 1H-pyrazolo[3,4-c]pyridine ring, a quinolinering, an isoquinoline ring, a 1,8-naphthyridine ring, an indoline ringand the like. The “9- or 10-membered bicyclic aromatic or partiallyunsaturated heterocycle containing, in the ring, one to threeheteroatoms independently selected from the group consisting of anitrogen atom and an oxygen atom” for Ring Q² is preferably anisoquinoline ring, an indazole ring, a benzimidazole ring, a1H-pyrrolo[2,3-c]pyridine ring, a 1H-pyrrolo[3,2-c]pyridine ring, afuro[3,2-b]pyridine ring, a 1H-pyrazolo[3,4-c]pyridine ring or anindoline ring.

In the present invention, the “5- to 8-membered saturated heterocyclecontaining, in the ring, one or two heteroatoms independently selectedfrom the group consisting of an oxygen atom and a nitrogen atom” refersto a 5- to 8-membered monocyclic saturated ring containing one or twoheteroatoms (a nitrogen atom or an oxygen atom) as a ring-constitutingatom, besides a carbon atom. Examples thereof include a pyrrolidinering, a piperidine ring, a piperazine ring, a morpholine ring, anazepane ring, an oxazepane ring, a 1,4-oxazepane ring, a 1,4-diazocanering and the like. The “5- to 8-membered saturated heterocyclecontaining, in the ring, one or two heteroatoms independently selectedfrom the group consisting of an oxygen atom and a nitrogen atom” forRing Q² is preferably a pyrrolidine ring, a piperidine ring, amorpholine ring or an azepane ring.

In the present invention, the “4- to 8-membered saturated heterocyclecontaining one nitrogen atom or one oxygen atom in the ring” refers to a4- to 8-membered monocyclic saturated ring containing one heteroatom (anitrogen atom or an oxygen atom) as a ring-constituting atom, besides acarbon atom. Examples thereof include an azetidine ring, an oxetanering, a pyrrolidine ring, a tetrahydrofuran ring, a piperidine ring, atetrahydropyran ring, an azepane ring and the like. The “4- to8-membered saturated heterocycle containing one nitrogen atom or oneoxygen atom in the ring” for Ring Q³ is preferably an azetidine ring, atetrahydropyran ring or a piperidine ring.

In the present invention, the “heterocycle containing a nitrogen atom inthe ring” refers to a heterocycle containing a nitrogen atom as aring-constituting atom, besides a carbon atom. Examples thereof includea piperidine ring, an azepane ring and the like.

In the present invention, the borono group refers to a group in whichtwo hydrogen atoms of a boranyl group are both substituted with hydroxygroups.

In the present invention, the dialkoxyboranyl group refers to a group inwhich two hydrogen atoms of a boranyl group are both substituted withalkoxy groups (e.g., a methoxy group, an ethoxy group, etc.). Examplesthereof include a dimethoxyboranyl group, a diethoxyboranyl group andthe like.

In the present invention, the dioxaborolanyl group refers to a groupderived from a ring formed by two alkoxy groups bonded to the boron atomof the above dialkoxyboranyl group taken together with the boron atom.Examples thereof include a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-ylgroup and the like.

In the present invention, menin refers to a tumor-suppressor proteinidentified as a causal factor of multiple endocrine neoplasia type 1(MEN1), which is an ubiquitously expressed nucleoprotein that isinvolved in DNA processing, modified proteins, protein-modifyingchromatin, and interactions with numerous transcription factors(Agarwal, et al.; Horm Metab Res, 2005, 37(6):369-374).

In the present invention, the MLL protein refers to MLL1, MLL2, a MLLfusion protein or a MLL partial tandem duplication protein.

In the present invention, the MLL1 refers to MLL1 (also known as KMT2A)protein, which is one of methyltransferases belonging to MLL (mixedlineage leukemia) family. In the present invention, the term MLL1 geneindicates a gene encoding the protein.

In the present invention, the MLL2 refers to MLL2 (also known as KMT2D)protein, which is one of methyltransferases belonging to MLL (mixedlineage leukemia) family. In the present invention, the term MLL2 geneindicates a gene encoding the protein.

In the present invention, the MLL fusion protein refers to a chimericprotein produced by transcription and expression of a chimeric genecaused by chromosomal translocation of a MLL gene.

In the present invention, the MLL partial tandem duplication (PTD)protein refers to an abnormal protein produced by transcription andexpression of an abnormal gene caused by chromosomal duplication of aMLL gene.

In the present invention, the interaction between menin and one or moreproteins selected from the group consisting of MLL1, MLL2, a MLL fusionprotein and a MLL partial tandem duplication protein refers to aninteraction between protein molecules formed by menin and MLL1, MLL2, aMLL fusion proteins or a MLL partial tandem duplication protein. Whentwo or more types of MLL proteins are present in the same system, two ormore interactions between protein molecules formed independently bymenin and each MLL protein may coexist.

In the present invention, the terms “tumor” and “cancer” are usedinterchangeably. Furthermore, in the present invention, tumor, malignanttumor, cancer, malignant neoplasm, carcinoma, sarcoma and the like maybe collectively referred to as “tumor” or “cancer”. Moreover, the terms“tumor” and “cancer” also include pathological conditions categorizedinto a premalignant stage in some cases, such as myelodysplasticsyndrome.

As used herein, the term “treat” and its derivatives mean remission,alleviation or delay of exacerbation of clinical symptoms of diseases,illnesses, disorders and the like (hereinafter referred to as “diseasesand the like”) in a patient who develops the diseases and the like.

As used herein, the term “prevent” and its derivatives mean inhibiting,suppressing, controlling, slowing or stopping the onset of clinicalsymptoms of the diseases and the like in a mammal who may develop thediseases and the like, but have not yet developed, or are concernedabout recurrence of the diseases and the like after treatment.

In the present invention, the “Bcl-2 inhibitor” refers to a drug thatbinds to Bcl-2, which is a protein having an anti-apoptotic action, toinhibit the anti-apoptotic action, and as a result, induces apoptosis toexert an anti-cancer action. In the present invention, the “Bcl-2inhibitor” is preferably Venetoclax.

In the present invention, the “pyrimidine antimetabolite” refers to adrug that has a partial structure similar to that of a pyrimidine base,and inhibits nucleic acid biosynthesis to prevent the growth anddivision of tumor cells, and as a result, exerts an anti-cancer action.

In the present invention, the “pyrimidine antimetabolite” is preferablyCytarabine.

In the present invention, the “DNA methyltransferase inhibitor” refersto a drug that inhibits an enzyme that catalyzes transmethylation ofDNA, and as a result, exerts an anti-cancer action. In the presentinvention, the “DNA methyltransferase inhibitor” is preferablyAzacitidine.

In the present invention, Venetoclax is4-(4-{[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-[(3-nitro-4-{[(oxan-4-yl)methyl]amino}phenyl)sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide(CAS registry number: 1257044-40-8), and is also referred to asVENCLEXTA (registered trademark), VENCLYXTO (registered trademark) orVenetoclax. It is readily available as a commercial product.

In the present invention, Azacitidine is4-amino-1-β-D-ribofuranosyl-1,3,5-triazin-2(1H)-one (CAS registrynumber: 320-67-2), and is also referred to as 5-Azacitidine or 5Aza, oras Vidaza (registered trademark). It is readily available as acommercial product.

In the present invention, Cytarabine is 1-β-D-arabinofuranosylcytosine(CAS registry number: 147-94-4), and is also referred to as Ara-C orAraC, or as CYLOCIDE (registered trademark). It is readily available asa commercial product.

In the present invention, Venetoclax, Azacitidine or Cytarabine may be afree form, a solvate, any of various pharmaceutically acceptable salts,or in the form of a pharmaceutical composition contained with variouspharmaceutically acceptable carriers, and the like.

In the present invention, the term “administered in combination” meansthat both drugs are taken into the body of the subject to beadministered for a certain period of time. Both drugs may beadministered in a single formulation, or each may be formulatedseparately and administered separately. When they are formulatedseparately, the timing of their administrations is not particularlylimited, and they may be administered at the same time, at differenttimes at intervals, or on different days.

When they are administered at different times or on different days, theorder of their administrations is not particularly limited. Generally,their formulations are administered according to their respectiveadministration methods, so that the formulations may be administered inthe same number of doses or in a different number of doses. In addition,when they are formulated separately, the respective administrationmethods (administration routes) of the formulations may be the same aseach other, or the formulations may be administered by differentadministration methods (administration routes). Moreover, both drugs donot have to exist in the body at the same time, and may be taken intothe body for a certain period of time (e.g., one month, preferably oneweek, more preferably several days, even more preferably one day). Oneof the active ingredients may have disappeared from the body at the timeof administration of the other active ingredient.

Suitable substituents in the compound of the present invention will bedescribed below.

R¹ is preferably a hydrogen atom or a methyl group. R¹ is morepreferably a hydrogen atom.

R² is preferably a hydrogen atom or a methyl group. R² is morepreferably a hydrogen atom.

One of R³ and R⁴ is preferably a hydrogen atom, a hydroxy group, afluorine atom, a methoxy group, a dimethylcarbamoyl group, or anoxazol-2-yl group, more preferably a hydrogen atom or a hydroxy group.The other of R³ and R⁴ is preferably a hydrogen atom, a hydroxy group, afluorine atom, or a methoxy group, more preferably a hydrogen atom or ahydroxy group.

The moiety represented by the following formula (5) in the formula (1)is preferably the following formula (5A) or (5B).

wherein * is bonded to the nitrogen atom to which R² is bonded, ** isbonded to the nitrogen atom to which R⁵ is bonded, R¹⁶ is a hydrogenatom, a halogen atom, a hydroxy group, a di(C₁₋₆ alkyl)carbamoyl group,an oxazol-2-yl group, or a C₁₋₆ alkoxy group, R¹⁷ is a hydrogen atom ora halogen atom, and R¹⁸ is a C₁₋₆ alkoxy group.

The moiety represented by the following formula (5) in the formula (1)is more preferably any of the following formulas (6A) to (6D), stillmore preferably (6A) or (6B).

wherein * is bonded to the nitrogen atom to which R² is bonded, ** isbonded to the nitrogen atom to which R⁵ is bonded, and R¹⁹ is a hydrogenatom, a hydroxy group, a dimethylcarbamoyl group, an oxazol-2-yl group,or a methoxy group.

The moiety represented by the following formula (5) in the formula (1)is preferably the following formula (7A).

wherein * is bonded to the nitrogen atom to which R² is bonded, ** isbonded to the nitrogen atom to which R⁵ is bonded, R²⁰ is a hydrogenatom or a hydroxy group, and R²¹ is a hydrogen atom, a hydroxy group, ora C₁₋₆ alkoxy group.

The moiety represented by the following formula (5) in the formula (1)is more preferably any of the following formulas (8A) to (8F), stillmore preferably any of (8A) to (8E).

wherein * is bonded to the nitrogen atom to which R² is bonded, ** isbonded to the nitrogen atom to which R⁵ is bonded, R²² is a hydrogenatom, a hydroxy group or a methoxy group, R²³ is a hydroxy group or amethoxy group, and R²⁴ is a hydrogen atom or a hydroxy group.

The moiety represented by the following formula (5) in the formula (1)is most preferably any of the following formulas (9A) to (9C). Among thefollowing formulas (9A) to (9C), it is preferably (9B) or (9C), morepreferably (9B).

wherein * is bonded to the nitrogen atom to which R² is bonded, and **is bonded to the nitrogen atom to which R⁵ is bonded.

R⁵ is preferably a hydrogen atom, a methyl group, an ethyl group, or a2-hydroxyethyl group. R⁵ is more preferably a methyl group.

R⁶ is preferably a hydrogen atom, a methyl group, a chlorine atom, amethoxy group, an amino group, or a methylamino group. R⁶ is morepreferably a hydrogen atom, a chlorine atom, a methoxy group, an aminogroup, or a methylamino group. R⁶ is still more preferably a hydrogenatom, a chlorine atom, a methoxy group, or a methylamino group.

As to R⁷ and R⁸, preferably, R⁷ and R⁸ are taken together with thecarbon atom to which R⁷ is bonded and the carbon atom to which R⁸ isbonded to form the following formula (2A) or (2B).

wherein the dotted circle indicates that the ring is aromatic, thecarbon atom marked with a is the carbon atom to which R⁸ is bonded, thecarbon atom marked with b is the carbon atom to which R⁷ is bonded, X isCH or a nitrogen atom, and R⁹ is a halogeno C₁₋₆ alkyl group, a C₃₋₈cycloalkyl group, a C₃₋₈ cycloalkyl C₁₋₆ alkyl group, a C₁₋₆ alkoxy C₁₋₆alkyl group, or an oxetanyl group.

R⁹ is preferably a 2,2,2-trifluoroethyl group, a cyclopropyl group, acyclopropylmethyl group, a methoxymethyl group, or an oxetan-3-yl group.R⁹ is more preferably a 2,2,2-trifluoroethyl group, a cyclopropyl group,or a cyclopropylmethyl group. R⁹ is still more preferably a2,2,2-trifluoroethyl group.

As to R⁷ and R⁸, more preferably, R⁷ and R⁸ are taken together with thecarbon atom to which R⁷ is bonded and the carbon atom to which R⁸ isbonded to form form the following formula (10A).

wherein the dotted circle indicates that the ring is aromatic, thecarbon atom marked with a is the carbon atom to which R⁸ is bonded, andthe carbon atom marked with b is the carbon atom to which R⁷ is bonded.

As to R⁷ and R⁸, preferably, R⁷ is a hydrogen atom, and R⁸ is thefollowing formula (3).

wherein * indicates a bonding site, R¹⁰ is a di(C₁₋₆ alkyl)carbamoylgroup, a (C₁₋₆ alkyl)pyrimidinyl group, a (C₁₋₆ alkyl)phenyl group, or a(C₁₋₆ alkyl)pyrazolyl group, R¹¹ is a hydrogen atom or a halogen atom,and R¹² is a halogen atom.

R¹⁰ is preferably a diisopropylcarbamoyl group, a4-isopropylpyrimidin-5-yl group, a 2-isopropylphenyl group, or a1-isopropylpyrazol-5-yl group. R¹¹ is preferably a hydrogen atom or afluorine atom. R¹² is preferably a fluorine atom.

As to R⁷ and R⁸, more preferably R⁷ is a hydrogen atom, and R⁸ is thefollowing formula (11A) or (11B).

wherein * indicates a bonding site, R²⁵ is a diisopropylcarbamoyl group,a 4-isopropylpyrimidin-5-yl group, a 2-isopropylphenyl group, or a1-isopropylpyrazol-5-yl group, and R²⁶ is a diisopropylcarbamoyl group.

m is preferably 1.

n is preferably 1.

Ring Q¹ is preferably any of the following (i) to (vii).

(i) a benzene ring optionally having one or two substituentsindependently selected from the above Group A;(ii) a pyridine ring optionally having one or two substituentsindependently selected from the above Group A;(iii) a 1,3-thiazole ring or a pyrazole ring (the 1,3-thiazole ring orpyrazole ring optionally has one substituent independently selected fromthe above Group A);(iv) a cyclohexane ring optionally having one substituent independentlyselected from the above Group A;(v) a cyclohexene ring optionally having one substituent independentlyselected from the above Group A;(vi) a piperidine ring optionally having one substituent independentlyselected from the above Group A; or(vii) an indole ring optionally has one or two substituentsindependently selected from the above Group B.

When m is 0, then Ring Q¹ is more preferably any of the following (i) to(iv).

(i) a benzene ring optionally having one or two substituentsindependently selected from the above Group A;(ii) a 1,3-thiazole ring or a pyrazole ring, each optionally having onesubstituent independently selected from the above Group A;(iii) a cyclohexane ring optionally having one substituent independentlyselected from the above Group A; or(iv) an indole ring optionally having one substituent independentlyselected from the above Group B.

When m is 0, then Ring Q¹ is still more preferably a phenyl group, a4-hydroxyphenyl group, a 4-[3-(prop-2-enoylamino)propylcarbamoyl]phenylgroup, a 4-[3-(vinylsulfonylamino)propylcarbamoyl]phenyl group, a3-fluoro-4-(2-hydroxyethoxy)phenyl group, a thiazol-5-yl group, acyclohexyl group, or a 2-cyano-1H-indol-5-yl group.

When m is 1, then Ring Q¹ is more preferably any of the following (i) to(vii).

(i) a benzene ring optionally having one substituent independentlyselected from the above Group A;(ii) a pyridine ring optionally having one substituent independentlyselected from the above Group A;(iii) a pyrazole ring optionally having one substituent independentlyselected from the above Group A;(iv) a cyclohexane ring optionally having one substituent independentlyselected from the above Group A;(v) a cyclohexene ring optionally having one substituent independentlyselected from the above Group A;(vi) a piperidine ring optionally having one substituent independentlyselected from the above Group A; or(vii) an indole ring optionally has one or two substituentsindependently selected from the above Group B.

When m is 1, then Ring Q¹ is still more preferably any of the followingformulas (12A) to (12H).

wherein * is bonded to Z, ** is bonded to the carbon atom to which R¹ isbonded, R²⁷ is a hydrogen atom, a halogen atom, a C₁₋₆ alkoxy group, ora C₁₋₆ alkyl group, J is a nitrogen atom or CR²⁹, R²⁹ is a halogen atom,and R²⁸ is a hydrogen atom or a C₁₋₆ alkyl group.

When m is 1, then Ring Q¹ is most preferably the following formula (13A)or (13B).

wherein * is bonded to Z, ** is bonded to the carbon atom to which R¹ isbonded, and R³⁰ is a hydrogen atom, a fluorine atom, a methyl group, ora methoxy group.

Ring Q² is preferably any of the following (i) to (vii).

(i) a benzene ring optionally having one to three substituentsindependently selected from the above Group C;(ii) a pyridine ring optionally having one to three substituentsindependently selected from the above Group C;(iii) a pyridazine ring, a pyrazine ring or a pyrimidine ring (thepyridazine ring, pyrazine ring or pyrimidine ring optionally has one tothree substituents independently selected from the above Group C);(iv) a pyrazole ring, an imidazole ring, a 1,3-thiazole ring, a1,3-oxazole ring or a 4H-1,2,4-triazole ring (the pyrazole ring,imidazole ring, 1,3-thiazole ring, 1,3-oxazole ring or 4H-1,2,4-triazolering optionally has one substituent independently selected from theabove Group C);(v) an isoquinoline ring, an indazole ring, a benzimidazole ring, a1H-pyrrolo[2,3-c]pyridine ring, a 1H-pyrrolo[3,2-c]pyridine ring, afuro[3,2-b]pyridine ring, a 1H-pyrazolo[3,4-c]pyridine ring or anindoline ring (the isoquinoline ring, indazole ring, benzimidazole ring,1H-pyrrolo[2,3-c]pyridine ring, 1H-pyrrolo[3,2-c]pyridine ring,furo[3,2-b]pyridine ring, 1H-pyrazolo[3,4-c]pyridine ring or indolinering optionally has one or two substituents independently selected fromthe above Group D);(vi) a pyrrolidine ring, a piperidine ring, a morpholine ring or anazepane ring (the pyrrolidine ring, piperidine ring, morpholine ring orazepane ring optionally has one substituent independently selected fromthe above Group E); or(vii) a cyclohexane ring optionally having one substituent independentlyselected from the above Group E.

When W is the above formula (4A), then Ring Q² is more preferably any ofthe following formulas (14A) to (14F).

wherein * indicates a bonding site, T is CH or a nitrogen atom, R³¹ is ahydrogen atom, a C₁₋₆ alkoxy group, a halogeno C₁₋₆ alkoxy group, or a(²H₃)methoxy group, R³² is a hydrogen atom, a C₁₋₆ alkyl group, ahalogen atom, a C₁₋₆ alkoxy group, a cyano group, a di(C₁₋₆ alkyl)aminogroup, a halogeno C₁₋₆ alkyl group, a C₁₋₆ alkylamino group, a C₁₋₆alkylsulfonyl group, a C₁₋₆ alkoxy C₁₋₆ alkoxy group, a halogeno C₁₋₆alkoxy group, a hydroxy C₁₋₆ alkyl group, a C₁₋₆ alkyl (2-C₃₋₆alkenoyl)amino group, a (²H₃)methoxy group, or a bis [(²H₃)methyl]aminogroup, or R³ 1 and R³² are taken together to form an ethylenedioxygroup, R³³ and R³⁵ are each independently a hydrogen atom, a halogenatom, a C₁₋₆ alkoxy group, a C₁₋₆ alkyl(C₁₋₆ alkylsulfonyl)amino group,a (C₁₋₆ alkyl)carbamoyl group, a di(C₁₋₆ alkyl)sulfamoyl group, a 2-C₃₋₆alkenoylamino group, or a C₁₋₆ alkylsulfonyl C₁₋₆ alkyl group, R³⁴ is ahydrogen atom or a halogen atom, R³⁶ is a halogen atom, R³⁷ is a C₁₋₆alkoxy group, R³⁸ is a halogen atom, R³⁹ is a C₁₋₆ alkyl group or a C₁₋₆alkylsulfonyl group, R⁴⁰ is a C₁₋₆ alkyl group or a C₁₋₆ alkylsulfonylgroup, U1 is CH or a nitrogen atom, U² is CR⁴¹ or a nitrogen atom, andR⁴ 1 is a hydrogen atom or a halogen atom.

R³¹ is preferably a hydrogen atom, a methoxy group, a difluoromethoxygroup, or a (²H₃)methoxy group.

R³² is preferably a hydrogen atom, a methyl group, a fluorine atom, achlorine atom, a methoxy group, a cyano group, a dimethylamino group, atrifluoromethyl group, a methylamino group, a methylsulfonyl group, amethoxyethoxy group, a difluoromethoxy group, a hydroxymethyl group, amethyl(prop-2-enoyl)amino group, a (²H₃)methoxy group, or abis[(²H₃)methyl]amino group.

R³³ and R³⁵ are each independently preferably a hydrogen atom, afluorine atom, a methoxy group, a prop-2-enoylamino group, amethyl(methylsulfonyl)amino group, a methylcarbamoyl group, adimethylsulfamoyl group, or a methylsulfonylmethyl group.

R³⁴ is preferably a hydrogen atom or a fluorine atom.

R³⁶ is preferably a fluorine atom.

R³⁷ is preferably a methoxy group.

R³⁸ is preferably a fluorine atom.

R³⁹ is preferably a methyl group or a methylsulfonyl group.

R⁴⁰ is preferably a methyl group or a methylsulfonyl group.

R⁴¹ is preferably a hydrogen atom or a fluorine atom.

When W is the above formula (4A), then Ring Q² is more preferably a5,6-dimethoxypyrazin-2-yl group, a 4,5-dimethoxypyrimidin-2-yl group, a4-pyridyl group, a 2,4-difluoro-3-methoxy-phenyl group, a4,5-dimethoxy-2-pyridyl group, a morpholino group, an oxazol-2-yl group,a 4H-1,2,4-triazol-3-yl group, a 5-oxopyrrolidin-2-yl group, a2-oxopyrrolidin-1-yl group, a cyclohexyl group, a 2-methoxythiazol-5-ylgroup, a furo[3,2-b]pyridin-6-yl group, an indolin-1-yl group, a3-hydroxy-1-piperidyl group, an azepan-1-yl group, a4-chloro-1H-pyrrolo[3,2-c]pyridin-7-yl group, a1-methylpyrazolo[3,4-c]pyridin-4-yl group, a benzimidazol-1-yl group, a4-isoquinolyl group, a 1-(difluoromethyl)-4-methoxy-6-oxo-pyridazin-3-ylgroup, or a 6-oxo-1H-pyridin-3-yl group.

When W is the above formula (4A), then Ring Q² is still more preferablyany of the following formulas (15A) to (15C).

wherein * indicates a bonding site, R⁴² is a methyl group, a chlorineatom, a methoxy group, a cyano group, a dimethylamino group, or a bis[(²H₃)methyl]amino group, R⁴³ is a methoxy group or a (²H₃)methoxygroup, and R⁴⁴ is a chlorine atom, a methoxy group, a methoxyethoxygroup, a dimethylamino group, a difluoromethoxy group, or a (²H₃)methoxygroup.

When W is the above formula (4A), then Ring Q² is most preferably any ofthe following formulas (16A) to (16G).

wherein * indicates a bonding site.

When W is the above formula (4B), then Ring Q² is more preferably thefollowing formula (17A) or (17B).

wherein * is bonded to Y, and ** is bonded to Z.

When W is the above formula (4B), then Ring Q³ is preferably any of thefollowing formulas (18A) to (18D).

wherein * indicates a bonding site, R⁴⁵ is a hydrogen atom or a halogenatom, R⁴⁶ is a C₁₋₆ alkylsulfonyl group, and V is a nitrogen atom or CH.

When W is the above formula (4B), then Ring Q³ is more preferably aphenyl group, an azetidin-1-yl group, a 3-pyridyl group, a6-chloro-3-pyridyl group, a tetrahydropyran-3-yl group, or a1-methylsulfonyl-4-piperidyl group.

When W is the above formula (4B), then Y is preferably a single bond oran oxygen atom.

Z is preferably a single bond, —NH—, an oxygen atom, —SO₂—, —CH₂—,*—CH₂—NHC(═O)—**, *—CH₂CH₂—O—**, or *—CH₂—NH—**, wherein * is bonded toRing Q², and ** is bonded to Ring Q¹.

Z is more preferably a single bond.

W is preferably the above formula (4A).

The compound of the present invention is preferably one selected fromthe following compounds or pharmaceutically acceptable salts thereof(preferably hydrochloride, succinate, benzenesulfonate, maleate,fumarate, mucate, or adipate, more preferably succinate,benzenesulfonate, maleate, fumarate, mucate, or adipate):

-   5-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-3-methoxypyridine-2-carbonitrile,-   (1R,2S,4R)-4-[({4-[1-(methanesulfonyl)-1H-indazol-4-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(5-methoxy-6-methylpyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(6-chloro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(6-fluoro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(6-chloro-5-methoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   2-[(4-{[(1S,2R,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-5-fluoro-N,N-di(propan-2-yl)benzamide,-   (1R,2S,4R)-2-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)cyclopentan-1-ol,-   (1R,3S)—N³-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-N′-methyl-N′-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine,-   (1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   6-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-4-methoxypyridazine-3-carbonitrile,-   (1S,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-[({4-[5-methoxy-6-(2-methoxyethoxy)pyridazin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(4,5-dimethoxypyridin-2-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-[({4-[6-(difluoromethoxy)-5-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-{[(4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}phenyl)methyl]amino}-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,-   (1R,2S,4R)-4-({[4-(6-{bis[(²H₃)methyl]amino}-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,    and-   (1R,2S,4R)-4-{[(4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}phenyl)methyl]amino}-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol.

The compound of the present invention is more preferably one selectedfrom the following compounds or pharmaceutically acceptable saltsthereof (preferably hydrochloride, succinate, benzenesulfonate, maleate,fumarate, mucate, or adipate, more preferably succinate,benzenesulfonate, maleate, fumarate, mucate, or adipate):

-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol,    and-   (1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol.

The compound of the present invention or a pharmaceutically acceptablesalt thereof or the crystal of the present invention has excellentproperties in terms of menin-MLL inhibitory action, solubility, cellmembrane permeability, oral absorption, blood concentration, metabolicstability, tissue transferability, bioavailability, in vitro activity,in vivo activity, rapid onset of drug effect, sustainability of drugeffect, physical stability, drug interaction, toxicity, and the like,and is useful as a drug.

In one embodiment, the present invention relates to a pharmaceuticalcomposition comprising, as an active ingredient, the compoundrepresented by the general formula (1) or a pharmaceutically acceptablesalt thereof or the crystal of the present invention, for the treatmentand/or prophylaxis of diseases that can be treated and/or prevented byinhibiting the interaction between an MLL protein and menin.

In another embodiment, the present invention relates to a method fortreating and/or preventing diabetes, comprising administering thecompound represented by the general formula (1) or a pharmaceuticallyacceptable salt thereof or the crystal of the present invention.

In another embodiment, the present invention relates to a method fortreating and/or preventing cancer, comprising administering the compoundrepresented by the general formula (1) or a pharmaceutically acceptablesalt thereof or the crystal of the present invention.

The disease to be treated is not particularly limited as long as itdepends on the interaction between menin and an MLL protein, andexamples thereof include cancers and diabetes (preferably cancer).

The type of cancer to be treated is not particularly limited as long asit is confirmed to be sensitive to the compound of the presentinvention. Examples thereof include blood cancer, brain tumor (e.g.,pediatric glioma, etc.), head/neck region cancer, esophageal cancer,stomach cancer, appendix cancer, colon cancer, anus cancer, gallbladdercancer, bile duct cancer, pancreatic cancer, gastrointestinal stromaltumor, lung cancer, liver cancer (e.g., hepatoma, etc.), mesothelioma,thyroid gland cancer, renal cancer, prostate cancer, neuroendocrinetumor, melanoma, breast cancer, endometrial cancer, cervical cancer,ovarian cancer, osteosarcoma, soft tissue sarcoma, Kaposi's sarcoma,myosarcoma, bladder cancer and testicular cancer. Preferred are bloodcancer, prostate cancer, breast cancer, hepatoma and pediatric glioma,and more preferred is blood cancer.

Examples of the blood cancer include mixed lineage leukemia (MLL),MLL-related leukemia, MLL-associated leukemia, MLL-positive leukemia,MLL-induced leukemia, rearranged mixed lineage leukemia (MLL-r),leukemia associated with a MLL rearrangement (a rearrangement of the MLLgene, MLL-rearranged leukemias), MLL-amplified leukemias, MLL partialtandem duplication leukemias (MLL-PTD leukemias), other leukemia/bloodcancers associated with constant expression of HOX and MEIS1 genes,acute leukemia, chronic leukemia, indolent leukemia, lymphoblasticleukemia, lymphocytic leukemia, myeloid leukemia, myelogenous leukemia,childhood leukemia, acute lymphoblastic leukemia (ALL), acute myeloidleukemia (AML), acute granulocytic leukemia, acute nonlymphocyticleukemia, chronic lymphocytic leukemia (CLL), chronic myelogenousleukemia (CML), therapy related leukemia, myelodysplastic syndrome(MDS), myeloproliferative disease (MPD), myeloproliferative neoplasia(MPN), multiple myeloma, myelodysplasia, plasma cell neoplasm, cutaneousT-cell lymphoma, lymphoid neoplasm, AIDS-related lymphoma, mycosisfungoides (granuloma fungoides), Alibert-Bazin syndrome, SezarySyndrome, hairy cell leukemia (HCL), T-cell prolymphocytic leukemia(T-PLL), large granular lymphocytic leukemia, meningeal leukemia,Hodgkin's lymphoma, non Hodgkin's lymphoma (malignant lymphoma),Waldenstrom's macroglobulinemia and the like. More preferred are acutemyeloid leukemia (AML) and acute lymphoblastic leukemia (ALL).

p53 is one of important factors that suppress carcinogenesis, anddeletion or mutation of the p53 gene has been observed in about half ofhuman cancers. It is known that mutations in p53 may promote cancer(gain-of-function p53 mutation), and cell growth is inhibited byallowing the compound having a menin-MLL inhibitory action on a cancercell line expressing gain-of-function p53 mutation (Zhu et al., Nature,2015, 525, 206-211.). Since the compound of the present invention or apharmaceutically acceptable salt thereof has a menin-MLL inhibitoryaction, it is effective for the treatment and/or prophylaxis of cancerexpressing gain-of-function p53 mutation. Examples of the cancerexpressing gain-of-function p53 mutation include blood cancer, braintumor, head/neck region cancer, esophageal cancer, stomach cancer,appendix cancer, colon cancer, anus cancer, gallbladder cancer, bileduct cancer, pancreatic cancer, gastrointestinal stromal tumor, lungcancer, liver cancer, mesothelioma, thyroid gland cancer, renal cancer,prostate cancer, neuroendocrine tumor, melanoma, breast cancer,endometrial cancer, cervical cancer, ovarian cancer, osteosarcoma, softtissue sarcoma, Kaposi's sarcoma, myosarcoma, bladder cancer andtesticular cancer.

The interaction between menin and a MLL fusion protein is known to beessential for the expression of several downstream oncogenes (e.g.,leukemia-related genes such as HOX, MEIS1, MYC, etc.) (Borkin et al.,Cancer Cell, 2015, 27, 589-602.). Since the compound of the presentinvention or a pharmaceutically acceptable salt thereof has a menin-MLLinhibitory action, it is effective for leukemia exhibiting expressioncharacteristics of HOX gene, MEIS1 gene, MYC gene etc.

Since the compound of the present invention or a pharmaceuticallyacceptable salt thereof or the crystal of the present invention has amenin-MLL inhibitory action, it is preferably used for diseasesdependent on the interaction between menin and a MLL protein. Examplesof the diseases dependent on the interaction between menin and a MLLprotein include blood cancer, prostate cancer, breast cancer, hepatoma,pediatric glioma and diabetes (e.g., see the following documents: bloodcancer (A1, A2, A3, A4), myelodysplastic syndrome (A1, A3), prostatecancer (B), breast cancer (C1, C2, C3), hepatoma (D), pediatric glioma(E), diabetes (F1, F2, F3)).

-   A1, Yokoyama et al., Cell, 2005, 123, 207-218.-   A2, Borkin et al., Cancer Cell, 2015, 27, 589-602.-   A3, Cierpicki and Grembecka. Future Med Chem. 2014, 447-462.-   A4, Kuehn M W et al., Cancer Discovery, 2016, 1166-1181.-   B, Malik et al., Nat. Med., 2015, 21, 344-352.-   C1, Dreijerink et al., Cancer Res., 2006, 66, 4929-4935.-   C2, Imachi et al., Breast Cancer Res. Treat., 2010, 122, 395-407.-   C3, Zhu et al., Nature, 2015, 525, 206-211.-   D, Xu et al., Proc. Natl. Acad. Sci. USA., 2013, 110, 17480-17485.-   E, Fumato et al., Science, 2014, 346, 1529-1533.-   F1, Wu et al., Curr. Mol. Med., 2008, 8(8), 805-815.-   F2, Chamberlain et al., J. Clin. Invest., 2014, 124, 4093-4101.-   F3, Yang et al., Proc. Natl. Acad. Sci. USA., 2010, 107,    20358-20363.

In another embodiment, the present invention relates to a pharmaceuticalcomposition comprising one drug selected from the group consisting of aBcl-2 inhibitor, a DNA methyltransferase inhibitor and a pyrimidineantimetabolite, and the compound of the present invention or apharmaceutically acceptable salt thereof or the crystal of the presentinvention, which are administered in combination.

In another embodiment, the present invention relates to a method fortreating cancer, comprising administering the compound of the presentinvention or a pharmaceutically acceptable salt thereof or the crystalof the present invention in combination with one drug selected from thegroup consisting of a Bcl-2 inhibitor, a DNA methyltransferase inhibitorand a pyrimidine antimetabolite.

In another embodiment, the present invention relates to the compound ofthe present invention or a pharmaceutically acceptable salt thereof orthe crystal of the present invention, which is administered incombination with one drug selected from the group consisting of a Bcl-2inhibitor, a DNA methyltransferase inhibitor and a pyrimidineantimetabolite.

One drug selected from the group consisting of a Bcl-2 inhibitor, a DNAmethyltransferase inhibitor and a pyrimidine antimetabolite, and thecompound of the present invention or a pharmaceutically acceptable saltthereof or the crystal of the present invention may be separatelycomprised as active ingredients in different formulations, or may becomprised in a single formulation. When they are separately comprised asactive ingredients in different formulations, their formulations may beadministered at the same time or different times.

In another embodiment, the present invention relates to a compositionfor inducing differentiation of leukemia cells, comprising the compoundof the present invention or a pharmaceutically acceptable salt thereofor the crystal of the present invention.

In another embodiment, the present invention relates to a method forinducing differentiation of leukemia cells, comprising administering thecompound of the present invention or a pharmaceutically acceptable saltthereof or the crystal of the present invention.

In the compound of the present invention, depending on the type andcombination of substituents, geometric isomers such as cis-forms andtrans-forms, tautomers, or optical isomers such as l-forms and d-forms(e.g., enantiomers or diastereomers) when the compound of the presentinvention has an asymmetric carbon atom can be present. The compound ofthe present invention includes all of these isomers and mixtures thereofin any ratio, unless otherwise specified.

In the present invention, the pharmaceutically acceptable salt includesboth pharmaceutically acceptable acid addition salts andpharmaceutically acceptable base addition salts.

When the compound of the present invention has a basic group such as anamino group etc., a pharmacologically acceptable acid addition salt canbe generally formed. Examples of the acid addition salt includehydrohalides such as hydrofluoride, hydrochloride, hydrobromide,hydroiodide etc.; inorganic acid salts such as nitrate, perchlorate,sulfurate, phosphate etc.; lower alkanesulfonates such asmethanesulfonate, trifluoromethanesulfonate, ethanesulfonate etc.; arylsulfonates such as benzenesulfonate, p-toluenesulfonate etc.; organicacid salts such as acetate, malate, fumarate, succinate, citrate,tartrate, oxalate, maleic acid, mucic acid, adipate etc.; and amino acidsalts such as ornithate, glutamate, aspartate etc., and preferred arehydrohalides, aryl sulfonates and organic acid salts. The acid additionsalt of the compound of the present invention is preferablyhydrochloride, succinate, benzenesulfonate, maleate, fumarate, mucate oradipate, more preferably succinate, benzenesulfonate, maleate, fumarate,mucate or adipate.

The acid addition salt of the compound of the present invention includesacid additions salt that can be formed by combining the acid to be addedto the compound of the present invention with the compound of thepresent invention in an any ratio. For example, the hydrochlorideincludes formable salts such as monohydrochloride, dihydrochloride,trihydrochloride etc., the fumarate includes formable salts such asmonofumarate, ½ fumarate etc., and the succinate includes formable saltssuch as monosuccinate, ⅔ succinate, ⅓ succinate etc.

When the compound of the present invention has an acidic group such as acarboxy group etc., a pharmacologically acceptable base addition saltcan be generally formed. Examples of the base addition salt includealkali metal salts such as sodium salt, potassium salt, lithium saltetc.; alkaline-earth metal salts such as calcium salt, magnesium saltetc.; inorganic salts such as ammonium salt etc.; and organic aminesalts such as dibenzylamine salt, morpholine salt, phenylglycinealkylester salt, ethylene diamine salt, N-methylglucamine salt, diethylaminesalt, triethylamine salt, cyclohexylamine salt, dicyclohexylamine salt,N,N′-dibenzylethylenediamine salt, diethanolamine salt,N-benzyl-N-(2-phenylethoxy)amine salt, piperazine salt,tetramethylammonium salt, tris(hydroxymethyl)aminomethane salt etc.

The compound of the present invention may be present as a non-solvate ora solvate. The solvate is not particularly limited as long as it ispharmacologically acceptable, and specifically, is preferably a hydrate,an ethanolate or the like. In addition, when a nitrogen atom is presentin the compound represented by the general formula (1), the compound maybe a N-oxide form. Such solvates and N-oxide forms are encompassedwithin the scope of the present invention. Moreover, the compound of thepresent invention can contain one or more isotopes at a non-naturalabundance, as a compound-constituting atom. Examples of the isotopeinclude deuterium (²H;D), tritium (³H; T), iodine-125 (¹²⁵I),carbon-14(¹⁴C) and the like. Furthermore, the compound of the presentinvention can be radiolabeled with a radioisotope such as tritium (³H),iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). The radiolabeled compound isuseful as a therapeutic or preventive agent, a research reagent (e.g.,an assay reagent), or a diagnostic agent (e.g., an in vivo imagediagnostic agent). The compound of the present invention containing anyradioactive or non-radioactive isotope in any ratio is encompassedwithin the scope of the present invention.

It is known that a low-molecular compound containing one or moredeuterium atoms (²H;D) as a compound-constituting hydrogen atom canexhibit a profile useful as a medicine (e.g., drug efficacy, safety,etc.) (Sanderson, Nature, 2009, DOI: 10.1038/458269a, Maltais et al, J.Med. Chem., 2009, 52, 7993-8001.). The compound of the present inventioninto which one or more deuterium atoms are introduced instead of thehydrogen atoms constituting the compound is also expected to exhibit thesame effect as above.

In the present invention, a crystal refers to a solid having an internalstructure formed by regularly three-dimensionally repeating constituentatoms or molecules, and is distinguished from an amorphous solid oramorphous substance not having such a regular internal structure. It canbe confirmed by employing powder X-ray crystal analysis or the like thatthe compound of the present invention or a salt thereof is in acrystalline state. In general, a peak value in powder X-ray diffractionmay inherently vary due to a difference in the measurement apparatus,sample or sample preparation, and hence the diffraction angle (2θ) canbe varied in a range of about ±0.2 (degrees). Therefore, it isunderstood that the value of the diffraction angle of the presentinvention encompasses numerical values falling in a range of about ±0.2.Accordingly, the scope of the present invention encompasses not onlycrystals having exactly the same diffraction angle (2θ), but alsocrystals having the same diffraction angle within the range of ±0.2, inpowder X-ray diffraction. Herein, the unit of the diffraction angle (2θ)is degree (also referred to as “°”), and the unit may be omitted in thedescription of the numerical value of the diffraction angle (2θ).

In the present invention, the crystal includes a crystal of the compoundrepresented by the general formula (1), a hydrate crystal of thecompound represented by the general formula (1), a solvate crystal ofthe compound represented by the general formula (1), a crystal of apharmaceutically acceptable salt of the compound represented by thegeneral formula (1), a hydrate crystal of a pharmaceutically acceptablesalt of the compound represented by the general formula (1), and asolvate crystal of a pharmaceutically acceptable salt of the compoundrepresented by the general formula (1). The hydrate crystal of thepresent invention may be in the form of, for example, 0.1, 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7,1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1,3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5,4.6, 4.7, 4.8, 4.9 or 5.0 hydrate, and the hydrated water may beincrease or decrease depending on the humidity.

The crystal of the present invention (hereinafter, sometimes to bereferred to as “the crystal of Example 131 of the present invention”,“the crystal of Example 132 of the present invention”, “the crystal ofExample 133 of the present invention”, “the crystal of Example 134 ofthe present invention”, “the crystal of Example 135 of the presentinvention”, “the crystal of Example 136 of the present invention”, “thecrystal of Example 137 of the present invention”, “the crystal ofExample 138 of the present invention” or “the crystal of Example 139 ofthe present invention”) can be stably supplied as a crystal of activepharmaceutical ingredient used in the production of pharmaceuticals, andhas excellent hygroscopicity or stability. The differences in thesecrystal forms are particularly distinguished by powder X-raydiffraction.

The crystal of Example 131 of the present invention has peaks atdiffraction angles (2θ) of 4.66±0.2, 7.02±0.2, 14.10±0.2, 16.68±0.2,17.46±0.2, 18.68±0.2, 21.34±0.2, 24.52±0.2, 25.54±0.2 and 28.22±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 131 of the present invention is preferablymonosuccinate.

The crystal of Example 131 of the present invention is preferablynon-hydrate.

The crystal of Example 132 of the present invention has peaks atdiffraction angles (2θ) of 10.92±0.2, 11.70±0.2, 12.40±0.2, 15.00±0.2,17.38±0.2, 18.16±0.2, 22.18±0.2, 22.62±0.2, 23.86±0.2 and 24.20±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 132 of the present invention is preferablymonobenzenesulfonate.

The crystal of Example 132 of the present invention is preferablytrihydrate.

The crystal of Example 133 of the present invention has peaks atdiffraction angles (2θ) of 4.64±0.2, 7.02±0.2, 7.46±0.2, 11.14±0.2,14.04±0.2, 16.76±0.2, 18.54±0.2, 19.76±0.2, 21.26±0.2 and 22.62±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 133 of the present invention is preferablymonomaleate.

The crystal of Example 133 of the present invention is preferablynon-hydrate.

The crystal of Example 134 of the present invention has peaks atdiffraction angles (2θ) of 4.80±0.2, 7.94±0.2, 9.66±0.2, 11.56±0.2,14.56±0.2, 17.62±0.2, 18.14±0.2, 20.46±0.2, 21.36±0.2 and 24.46±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 134 of the present invention is preferablymonofumarate.

The crystal of Example 134 of the present invention is preferablytetrahydrate.

The crystal of Example 135 of the present invention has peaks atdiffraction angles (2θ) of 7.14±0.2, 8.76±0.2, 12.26±0.2, 14.30±0.2,17.52±0.2, 23.40±0.2, 24.40±0.2, 24.86±0.2, 25.34±0.2 and 25.90±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 135 of the present invention is preferablytrihydrate.

The crystal of Example 136 of the present invention has peaks atdiffraction angles (2θ) of 8.06±0.2, 12.22±0.2, 12.52±0.2, 15.14±0.2,17.54±0.2, 18.56±0.2, 20.08±0.2, 23.48±0.2, 24.28±0.2 and 25.00±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 136 of the present invention is preferablymonofumarate.

The crystal of Example 136 of the present invention is preferablydihydrate.

The crystal of Example 137 of the present invention has peaks atdiffraction angles (2θ) of 6.56±0.2, 9.44±0.2, 9.94±0.2, 13.20±0.2,18.22±0.2, 18.86±0.2, 19.60±0.2, 22.68±0.2, 25.10±0.2 and 28.70±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 137 of the present invention is preferablymonomucate.

The crystal of Example 137 of the present invention is preferablytrihydrate.

The crystal of Example 138 of the present invention has peaks atdiffraction angles (2θ) of 5.88±0.2, 6.20±0.2, 9.18±0.2, 10.34±0.2,12.50±0.2, 13.70±0.2, 15.66±0.2, 17.82±0.2, 18.48±0.2 and 22.16±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 138 of the present invention is preferablymonoadipate.

The crystal of Example 138 of the present invention is preferablytrihydrate.

The crystal of Example 139 of the present invention has peaks atdiffraction angles (2θ) of 4.60±0.2, 6.60±0.2, 7.74±0.2, 8.02±0.2,9.26±0.2, 11.16±0.2, 12.00±0.2, 12.44±0.2, 13.22±0.2 and 19.66±0.2 in apowder X-ray diffraction diagram obtained through irradiation withcopper Kα line (λ=1.54 angstroms).

The crystal of Example 139 of the present invention is preferablymonosuccinate.

The crystal of Example 139 of the present invention is preferably 2.5hydrate.

The present invention encompasses a compound which can be converted tothe compound represented by the general formula (1), which is an activeingredient of the pharmaceutical composition of the present invention,with a reaction due to an enzyme, gastric acid and the like under thephysiological condition in the living body, that is, a compound whichcan be converted to the compound represented by the general formula (1)by enzymatic oxidation, reduction, hydrolysis and the like; and acompound which can be converted to the compound represented by thegeneral formula (1) by hydrolysis and the like due to gastric acid andthe like, as a “pharmaceutically acceptable prodrug compound”.

When the compound represented by the general formula (1) contains anamino group, examples of the prodrug include a compound obtained bysubjecting the amino group to acylation, alkylation or phosphorylation(e.g., a compound obtained by subjecting the amino group toeicosanoylation, alanylation, pentylaminocarbonylation,(5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylation,tetrahydrofurylation, pyrrolidylmethylation, pivaloyloxymethylation ortert-butylation) and the like. When the compound represented by thegeneral formula (1) contains a hydroxy group, examples of the prodruginclude a compound obtained by subjecting the hydroxy group toacylation, alkylation, phosphorylation or boration (e.g., a compoundobtained by subjecting the hydroxy group to acetylation, palmitoylation,propanoylation, pivaloylation, succinylation, fumarylation, alanylationor dimethylaminomethylcarbonylation) and the like. When the compoundrepresented by the general formula (1) contains a carboxyl group,examples of the prodrug include a compound obtained by subjecting thecarboxyl group to esterification or amidation (e.g., a compound obtainedby subjecting the carboxyl group to ethyl esterification, phenylesterification, carboxymethyl esterification, dimethylaminomethylesterification, pivaloyloxymethyl esterification, ethoxycarbonyloxyethylesterification or methylamidation) and the like.

The prodrug of the present invention can be produced from the compoundrepresented by the general formula (1) according to a method known perse. The prodrug of the present invention also includes a compound whichcan be converted to the compound represented by the general formula (1)under physiological conditions as described in “IYAKUHIN no KAIHATSU(Development of Pharmaceuticals)”, Vol. 7, Design of Molecules, p.163-198, published by HIROKAWA SHOTEN (1990).

[Production Method]

Next, typical production methods for the compound represented by thegeneral formula (1) will be described. The compound of the presentinvention can be produced according to various production methods, andthe production methods shown below are merely examples, and the presentinvention should not be construed as limited to these.

The compound represented by the general formula (1), pharmaceuticallyacceptable salts thereof and synthetic intermediates thereof can beproduced by employing various known production methods, with utilizingcharacteristics based on the basic skeleton or the type of substituent.Examples of the known methods include methods described in “ORGANICFUNCTIONAL GROUP PREPARATIONS”, the 2nd edition, ACADEMIC PRESS, INC.,1989, “Comprehensive Organic Transformations”, the 2nd edition, VCHPublishers Inc., 1999, and the like.

In the production, depending on the type of the functional groupcontained in the compound, it may sometimes be effective in theproduction technology to protect the functional group with anappropriate protecting group at the stage of a raw material orintermediate, or to substitute with a group that can be easily convertedto the functional group.

Examples of the functional group include an amino group, a hydroxygroup, a formyl group, a carbonyl group, and carboxy group and the like,and examples of the protecting group include protecting groups describedin “Protective Groups in Organic Synthesis”, the 5th edition, Wiley,2014 written by P. G. Wuts.

The protecting group or the group that can be easily converted to thefunctional group may be appropriately selected in accordance with thereaction conditions of the production method employed for producing thecompound.

According to such a method, after introducing the group and carrying outthe reaction, the desired compound can be obtained by removing theprotecting group or converting the group to a desired group, ifnecessary.

The prodrug of the compound can be produced by introducing a specificgroup at the stage of a raw material or intermediate, or by subjectingthe obtained compound to introduction of the group, as in the case ofthe above-mentioned protecting group. The reaction for producing aprodrug can be carried out by employing conventional methods known tothose skilled in the art, such as esterification, amidation,dehydration, hydrogenation and the like.

The compound represented by the general formula (1) can be produced, forexample, according to the following Methods A to E. The syntheticintermediates used in Method A to Method E can be produced, for example,according to the following Methods F to Y.

When the compound serving as a reaction substrate in the reaction ineach step of the following Methods A to Y has a functional group orpartial structure that inhibits the desired reaction, such as an aminogroup, a hydroxy group, a formyl group, a carbonyl group, a carboxygroup, a heteroatom on a cyclic compound and the like, a protectinggroup may be introduced into it or an introduced protecting group may beremoved therefrom, appropriately if necessary. Such a protecting groupis not particularly limited as long as it is a commonly used protectinggroup, and may be, for example, the protecting group described in theabove-mentioned “Protective Groups in Organic Synthesis (5th edition,2014)”. The reactions for the introduction and removal of theseprotecting groups can be carried out according to the conventionalmethods described in the above document.

In each compound of Methods A to Y, depending on the type of thefunctional group contained in the compound, the functional group can besubstituted with a group that can be easily converted to the functionalgroup at the stage of a raw material or intermediate. The conversion tothe desired functional group can be performed at an appropriate stageaccording to a known method. Examples of the known method includemethods described in the above-mentioned documents “ORGANIC FUNCTIONALGROUP PREPARATIONS”, “Comprehensive Organic Transformations” and thelike.

In each compound in the following Methods A to Y is isolated andpurified in the form of a non-solvate, a salt or any of various solvatessuch as a hydrate. The salt can be produced according to a conventionalmethod. Examples of the salt include hydrochloride, sulfurate and thelike; organic amine salts; and sodium salt, potassium salt and the like.

The solvent used in the reaction in each step of the following Methods Ato Y is not particularly limited as long as it does not inhibit thereaction but partially dissolves a starting material, and is selected,for example, from the following solvent group. The solvent groupincludes aliphatic hydrocarbons such as n-hexane, n-pentane, petroleumether and cyclohexane; aromatic hydrocarbons such as benzene, tolueneand xylene; halogenated hydrocarbons such as dichloromethane (methylenechloride), chloroform, carbon tetrachloride, dichloroethane,chlorobenzene and dichlorobenzene; ethers such as diethyl ether,diisopropyl ether, tetrahydrofuran (THF), dioxane, dimethoxyethane anddiethylene glycol dimethyl ether; ketones such as acetone, methyl ethylketone, methylisobutylketone and cyclohexanone; esters such as ethylacetate, propyl acetate, butyl acetate; nitriles such as acetonitrile,propionitrile, butyronitrile and isobutyronitrile; carboxylic acids suchas acetic acid and propionic acid; alcohols such as methanol, ethanol,1-propanol, 2-propanol, 1-butanol, 2-butanol, 2-methyl-1-propanol and2-methyl-2-propanol; amides such as formamide, N,N-dimethylformamide(DMF), N,N-dimethylacetamide, N-methyl-2-pyrrolidone,hexamethylphosphortriamide; sulfoxides such as dimethyl sulfoxide (DMSO)and tetrahydrothiophene 1,1-dioxide; water; and mixture thereof.

The acid used in the reaction in each step of the following Methods A toY is not particularly limited as long as it does not inhibit thereaction, and is selected from the following acid group. The acid groupincludes inorganic acids such as hydrochloric acid, hydrobromic acid,hydroiodic acid, phosphoric acid, sulfuric acid and nitric acid; organicacids such as acetic acid, propionic acid, trifluoroacetic acid andpentafluoropropionic acid; organic sulfonic acids such asmethanesulfonic acid, trifluoromethanesulfonic acid, p-toluenesulfonicacid and camphorsulfonic acid; and Lewis acids such as boron tribromide,indium(III) bromide, boron trifluoride, aluminium(III) chloride andtrimethylsilyl trifluoromethanesulfonate.

The base used in the reaction in each step of the following Methods A toY is not particularly limited as long as it does not inhibit thereaction, and is selected from the following base group. The base groupincludes alkali metal carbonates such as lithium carbonate, sodiumcarbonate, potassium carbonate and cesium carbonate; alkali metalhydrogencarbonates such as lithium hydrogencarbonate, sodiumhydrogencarbonate and potassium hydrogencarbonate; alkali metalhydroxides such as lithium hydroxide, sodium hydroxide and potassiumhydroxide; alkaline-earth metal hydroxides such as calcium hydroxide andbarium hydroxide; alkali metal hydrides such as lithium hydride, sodiumhydride and potassium hydride; alkali metal amides such as lithiumamide, sodium amide and potassium amide; alkali metal alkoxides such aslithium methoxide, sodium methoxide, sodium ethoxide, sodiumtert-butoxide and potassium tert-butoxide; lithium alkylamides such aslithium diisopropylamide; silylamides such as lithiumbistrimethylsilylamide and sodium bistrimethylsilylamide; alkyl lithiumssuch as n-butyllithium, sec-butyllithium and tert-butyllithium;alkylmagnesium halides such as methylmagnesium chlorides(methylmagnesium chloride), methylmagnesium bromides (methylmagnesiumbromide), methylmagnesium iodides (methylmagnesium iodide),ethylmagnesium chlorides (ethylmagnesium chloride), ethylmagnesiumbromides (ethylmagnesium bromide), isopropylmagnesium chlorides(isopropylmagnesium chloride), isopropylmagnesium bromides(isopropylmagnesium bromide) and isobutylmagnesium chlorides(isobutylmagnesium chloride); and organic amines such as triethylamine(TEA), tributylamine, N,N-diisopropylethylamine (DIPEA),1-methylpiperidine, 4-methylmorpholine, 4-ethylmorpholine, pyridine,picoline, 4-dimethylaminopyridine, 4-pyrrolidinopyridine,2,6-di-tert-butyl-4-methylpyridine, quinoline, N,N-dimethylaniline,N,N-diethylaniline, 1,5-diazabicyclo[4,3,0]-5-nonene (DBN),1,4-diazabicyclo[2,2,2]octane (DABCO),1,8-diazabicyclo[5,4,0]-7-undecene(DBU) and imidazole.

In the reaction in each step of the following Methods A to Y, thereaction temperature depends on the solvent, starting material, reagentsand the like, and the reaction time depends on the solvent, startingmaterial, reagents, reaction temperature and the like.

In the reaction in each step of the following Methods A to Y, the targetcompound of each step is isolated from the reaction mixture by aconventional method after completing the reaction. The target compoundis obtained, for example, by (i) filtering off an insoluble substancesuch as a catalyst etc., if necessary, (ii) extracting the targetcompound by adding water and a solvent immiscible with water (e.g.,dichloromethane, diethyl ether, ethyl acetate etc.) to the reactionmixture, (iii) washing the organic layer with water and drying theresultant with a desiccant such as anhydrous calcium sulfate etc., and(iv) evaporating the solvent. The obtained target compound can befurther purified, if necessary, by a conventional method, for example,recrystallization, reprecipitation, distillation or columnchromatography (including normal phase chromatography and reverse phasechromatography) using silica gel, alumina or the like. The obtainedtarget compound is identified by standard analysis techniques such aselemental analysis, NMR, mass spectroscopy, IR analysis etc., and itscomposition or purity can be thus analyzed. Alternatively, the targetcompound obtained in each step can be used directly in the next reactionwithout purification.

In each step of the following Methods A to Y, an optical isomer can beseparated and purified by fractional recrystallization using anoptically active amine such as (R)-(+)- or (S)-(−)-1-phenethylamineetc., or an optically active carboxylic acid such as (+)- or(−)-10-camphorsulfonic acid etc., or by separation using an opticallyactive column.

The deuterium (²H;D) substitute of the compound represented by thegeneral formula (1) can be produced, for example, by employing a methodcommonly used by those skilled in the art at an appropriate stage duringthe following Methods A to Y. Examples of the method generally used bythose skilled in the art include the methods described in Nature, 2007,446, 526-529., Angew. Chem. Int. Ed., 2007, 46, 7744-7765., J. Med.Chem., 2009, 52, 7993-8001., and the like.

The raw materials and reagents used in Methods A to Y employed for theproduction of the compound of the present invention may be a knowncompound, or can be produced from a known compound as a startingmaterial according to a known method or a method analogous thereto. Thestarting material known compound can also be purchased from commercialsuppliers.

Abbreviations Used Herein

Boc: tert-butoxycarbonylCbz: benzyloxycarbonylAlloc: allyloxycarbonylNs: 2-nitrobenzenesulfonyl (nosyl)MOM: methoxymethylTMS: trimethylsilylOTf: trifluoromethylsulfonyloxyTr: triphenylmethylPMB: p-methoxybenzylBOP: (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphateHATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphateCOMU:N-[1-(cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino(morpholino)]uroniumhexafluorophosphateEDC: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochlorideHOBt: 1-hydroxybenzotriazoleDPPA: diphenylphosphoryl azide

The compound represented by the general formula (1) can be producedaccording to the methods shown below.

Method A

In the scheme, PG is a protecting group for an amino group, and examplesthereof include a Boc group, a Cbz group, a Ns group, an Alloc group andthe like. The protecting groups described in the above-mentioned“Protective Groups in Organic Synthesis”, and the like can also be used.LG is a leaving group, and examples thereof include a halogen atom, ap-toluenesulfonyl group and the like.

Step A-1 is a step of obtaining Intermediate III from Intermediate I andIntermediate II. This step can be performed by heating Intermediate Iand Intermediate II in the presence of a base (e.g., DIPEA, etc.), in asolvent inert to the reaction (e.g., isopropyl alcohol, etc.).

Step A-2 is a step of removing PG. When PG is a Cbz group, the step canbe performed by treating Intermediate III with an acid (e.g.,iodotrimethylsilane, etc.) in a solvent inert to the reaction (e.g.,acetonitrile, etc.). When PG is a Boc group, the step can be performedby treating Intermediate III with an acid (e.g., hydrochloric acid,etc.) in a solvent inert to the reaction (e.g., dichloromethane, etc.).When PG is a Ns group, the step can be performed by reactingIntermediate III with an thiol (e.g., isopropylbenzenethiol, etc.) and abase (e.g., cesium carbonate, etc.) in a solvent inert to the reaction(e.g., a mixed solvent of THF and methanol, etc.). In addition, themethods described in the above-mentioned “Protective Groups in OrganicSynthesis” can also be applied.

Step A-3 is a step of obtaining the compound represented by the generalformula (1) wherein R² is a hydrogen atom, from Intermediate IV andIntermediate V. This step can be performed by reacting Intermediate IVand Intermediate V with a reducing agent (e.g., sodiumtriacetoxyborohydride, sodium cyanoborohydride, etc.) in a solvent inertto the reaction (e.g., dichloromethane, dichloroethane, etc.). Acatalyst such as tetraisopropoxytitanium and the like can also be usedto promote the reaction.

Step A-4 is a step of converting R² of the compound represented by thegeneral formula (1) wherein R² is a hydrogen atom to a C₁₋₆ alkyl group.This step can be performed by reacting the compound with an alkylatingagent (e.g., methyl trifluoromethanesulfonate, etc.) and a base (e.g.,pyridine, etc.) in a solvent inert to the reaction (e.g.,dichloromethane, etc.).

Intermediate III can also be produced using Intermediate I andIntermediate II′ (Method B).

Method B

In the scheme, PG is as defined above.

Step B can be performed by reacting Intermediate I and Intermediate II′with a condensing agent (e.g., a BOP reagent, etc.) and a base (e.g.,DBU, etc.) in a solvent inert to the reaction (e.g., acetonitrile,etc.), and then heating the mixture.

The compound represented by the general formula (1) can also be producedfrom Intermediate IV and Intermediate V′ (Method C). Intermediate V′ canbe produced, for example, according to the methods described in CANCERCELL. 2015, 27, 589-602.

Method C

In the scheme, LG is as defined above.

Step C-1 is a step of obtaining the compound represented by the generalformula (1) wherein R² is a hydrogen atom, from Intermediate IV andIntermediate V′. This step can be performed by reacting Intermediate IVand Intermediate V′ with a base (e.g., potassium carbonate, etc.) in asolvent inert to the reaction (e.g., DMF, etc.). When a protecting groupis present in the structure of Intermediate V′, the deprotection canalso be performed under a suitable reaction condition (e.g., a method ofreacting with tin tetrachloride and the like, in a solvent such asacetonitrile, and the like, in the case of a Boc group as a protectinggroup) after Step C-1 to convert to the desired structure.

Step C-2 is a step of converting R² of the compound represented by thegeneral formula (1) wherein R² is a hydrogen atom to a C₁₋₆ alkyl group.This step can be performed in the same manner as in Step A-4. When aprotecting group is present in the structure of Intermediate V′, thedeprotection can also be performed under a suitable reaction condition(e.g., a method of reacting with tin tetrachloride and the like, in asolvent such asacetonitrile, and the like, in the case of a Boc group asa protecting group) after Step C-2 to convert to the desired structure.

Each step shown in the above Methods A to C does not necessarily have tobe performed in the same order as long as it does not affect thereaction substrate and the reaction product, and, for example, it may beperformed in the following order (Method D).

Method D

In the scheme, PG and LG are as defined above.

When the compound represented by the general formula (1) is representedby the following compound (1′), it can also be produced via thefollowing Intermediate VIII (Method E).

Method E

In the scheme, R^(e1) is a substituent that can be reacted in thebelow-mentioned cross coupling reaction (Step E-2), such as a halogenatom (e.g., bromine, iodine, etc.), a trifluoromethylsulfonyloxy group(OTf group) and the like. R^(e2) is a borono group, a dialkoxyboranylgroup (e.g., a dimethoxyboranyl group, etc.), a dioxaborolanyl group (a4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, etc.) or the like. Z¹is —NH— or a single bond.

Step E-1 is a step of producing Intermediate VIII from Intermediate IVand Compound 1e. This step can be performed in the same manner as inStep A-3.

Step E-2 is a step of obtaining the compound represented by the generalformula (1′) wherein R² is a hydrogen atom, from Intermediate VIII andany one of Compounds 2e and 3e. This step can be performed by heatingIntermediate VIII and any one of Compounds 2e and 3e under nitrogenatmosphere in the presence of a metal catalyst (e.g.,bis(triphenylphosphine)palladium dichloride, etc.) and a base (e.g.,potassium carbonate, etc.), in a solvent inert to the reaction (e.g., amixed solvent of dimethoxyethane and water, etc.).

Step E-3 is a step of converting R² of in the compound represented bythe general formula (1′) wherein R² is a hydrogen atom to a C₁₋₆ alkylgroup. This step can be performed in the same manner as in Step A-4.

The production method of each intermediate will be described below.

The production methods of Intermediate I and Intermediate I′ will bedescribed. The production methods shown below are merely examples andshould not be construed as limited to these.

In the scheme, PG is as defined above.

Intermediate I or Intermediate I′ can also be converted to each other,if necessary, by appropriately combining both steps of (a) introductionof a separate protecting group on the nitrogen atom and (b) removal ofan unnecessary protecting group. These steps are general conversionreactions of protecting groups, and can be performed, for example, byemploying the method described in the above-mentioned “Protective Groupsin Organic Synthesis”.

When Intermediate I is represented by the following Compound I-1, I-2,I-3 or I-4, it can be produced, for example, according to Method F or I.Starting Material if is known, or is produced using a known compound asa starting material according to a known method or a method analogousthereto. Known compounds can be purchased from commercial suppliers.Examples of known document include Tetrahedron Asymmetry. 2013, 24,651-656, Tetrahedron. 2004, 60, 717-728, Bioorg. Med. Chem. 2006, 14,2242-2252, Tetrahedron. 2017, 73, 1381-1388 and the like. Compound ifcan also be synthesized according to Method G.

Method F

In the scheme, R^(f1) and R^(f2) are each independently a C₁₋₆ alkylgroup or a protected hydroxy C₁₋₆ alkyl group (e.g., a2-[tert-butyl(dimethyl)silyl]oxyethyl group, etc.). PG and PG′ are eachindependently a protecting group different from each other introduced onthe nitrogen atom, and examples thereof include a Boc group, a Cbzgroup, an Alloc group and the like, and the protecting groups describedin the above-mentioned “Protective Groups in Organic Synthesis (the 5thedition, 2014)”, and the like can also be used.

Step F-1 is a step of synthesizing Compound 2f from Compound 1f. Thisstep can be performed by heating Compound if together with an azidizingagent (e.g., diphenylphosphoryl azide (DPPA), etc.), a base (e.g.,triethylamine, etc.) and an alcohol (e.g., benzyl alcohol, allylalcohol, etc.) in a solvent inert to the reaction (e.g., toluene, etc.).

Step F-2 is a step of synthesizing Intermediate I-1 or I-2 from Compound2f (if necessary, the protecting group to be removed may be either PG orPG). Both steps can be performed under the same conditions as in StepA-2.

When R⁵ of Intermediate I or I′ is R^(f1) or R^(f2), Intermediate I-3 orI-4 can be produced by performing Step F-2, followed by Steps F-3 toF-5.

Step F-3 is a step of synthesizing Compound 3f-1 from Intermediate I-1,or a step of synthesizing Compound 3f-2 from Intermediate I-2. Bothsteps can be performed, for example, by reacting Intermediate I-1 orIntermediate I-2 with a nosylating agent (e.g., 2-nitrobenzenesulfonylchloride, etc.) and a base (e.g., DIPEA, etc.) in a solvent inert to thereaction (e.g., dichloromethane, etc.).

Step F-4 is a step of synthesizing Compound 4f-1 from Compound 3f-1, ora step of synthesizing, Compound 4f-2 from Compound 3f-2. Both steps canbe performed, for example, by reacting Compound 3f-1 or Compound 3f-2with an alkylating agent (e.g., iodomethane, etc.) and a base (e.g.,potassium carbonate, etc.) in a solvent inert to the reaction (e.g.,DMF, etc.).

Step F-5 is a step of synthesizing Intermediate I-3 from Compound 4f-1,or a step of synthesizing Intermediate I-4 from Compound 4f-2. Bothsteps can be performed, for example, by reacting Compound 4f-1 orCompound 4f-2 with a thiol (e.g., isopropylbenzenethiol, etc.) and abase (e.g., cesium carbonate, etc.) in a solvent inert to the reaction(e.g., a mixed solvent of THF and methanol, etc.).

When a functional group having a protecting group is present in R³ orR⁴, the protecting group may be removed at the same time as Step F-2 isperformed.

Intermediates I-1 to I-4 can also be converted to the desired compounds,which can be used as Intermediate I or a starting material for othersteps, if necessary, by appropriately combining both steps of (a)introduction of a separate protecting group on the nitrogen atom and (b)removal of an unnecessary protecting group. These steps are generalconversion reactions of protecting groups, and can be performed, forexample, by employing the method described in the above-mentioned“Protective Groups in Organic Synthesis”.

When the compound represented by the general formula (1f) is representedby the following Compound 1f-1, it can be produced according to MethodG. The starting materials are known, or are produced using a knowncompound as a starting material according to a known method or a methodanalogous thereto.

Method G

In the scheme, PG′ is as defined above. R^(g1) is a protecting group fora carboxy group (e.g., a methyl group, an ethyl group, a MOM group,etc.). R^(g2) is a C₁₋₆ alkyl group (e.g., a methyl group, an ethylgroup, etc.) or a protecting group for a hydroxy group (e.g., MOM,etc.).

Step G-1 can be performed by reacting Compound 1g-1 or Compound 1g-2with an alkylating agent (e.g., chloromethyl methyl ether, etc.) and abase (e.g., DIPEA, etc.) in the presence of a reaction promoter (e.g.,sodium iodide, etc.), in a solvent inert to the reaction (e.g.,dimethoxyethane, etc.), and then heating the mixture. In this step,either Compound 1g-1 or Compound 1g-2 may be used as a raw material.While the reaction conditions of this step depend on the substrate, thisstep can also be performed by reacting Compound 1g-1 or Compound 1g-2with an alkylating agent (e.g., iodomethane, etc.) in the presence of ametal catalyst (e.g., silver(I) oxide) and an additive (e.g., molecularsieve, etc.), in a solvent inert to the reaction (e.g., dichloromethane,etc.), and then heating the mixture. The step of converting Compound1g-1 to Compound 2g-1 can also be performed in two steps: protection ofthe carboxy group and protection of the hydroxy group.

Step G-2 is a step of synthesizing Compound if-1 from Compound 2g-1 orCompound 2g-2. This step can be performed, for example, by treatingCompound 2g-1 or Compound 2g-2 with a base (e.g., aqueous sodiumhydroxide solution, etc.) in a solvent inert to the reaction (e.g., amixed solvent of methanol and THF, etc.).

Intermediate I′-1 can be produced as follows (Method H). The rawmaterial, Compound 1h can be synthesized, for example, according toMethod F.

Method H

In the scheme, PG and PG′ are as defined above. R^(h) is C1-20₆ alkyl.

Step H-1 can be performed in the same manner as in Step G-1.

Step H-2 can be performed in the same manner as in Step A-2.

When Intermediate I is represented by the following Compound I-5, it canalso be produced as follows (Method I). The raw material, Compound 1ican be synthesized, for example, according to Method G.

Method I

In the scheme, R^(i1) and R^(i2) are each independently a C₁₋₆ alkylgroup.

Step I-1 is a step of obtaining Compound 2i from Compound 1i. This stepcan be performed by reacting Compound 1i with Compound 7i in thepresence of a condensing agent (e.g., EDC, etc.), a catalyst (e.g.,HOBt, etc.) and abase (e.g., triethylamine etc.), in a solvent inert tothe reaction (e.g., dichloromethane, etc.)

Step I-2 is a step of obtaining Compound 3i from Compound 2i. This stepcan be performed by treating Compound 2i with an acid (e.g.,hydrochloric acid, etc.) in a solvent inert to the reaction (e.g.,1,4-dioxane, etc.).

Step I-3 is a step of obtaining Compound 4i from Compound 3i. Thisreaction can be performed by reacting Compound 3i with a carboxylic acid(e.g., 4-nitrobenzoic acid, etc.) in the presence of a phosphinecompound (e.g., triphenylphosphine, etc.) and an azodicarboxylatecompound (e.g., diisopropyl azodicarboxylate, etc.), in a solvent inertto the reaction (e.g., THF, etc.).

Step I-4 is a step of obtaining Compound 5i from Compound 4i. This stepcan be performed by treating Compound 4i with a base (e.g., potassiumcarbonate, etc.) in a solvent inert to the reaction (e.g., ethanol,etc.).

Step I-5 is a step of obtaining Compound 6i from Compound 5i. This stepcan be performed by reacting Compound 5i with diphenylphosphoryl azide(DPPA) in the presence of an azodicarboxylate compound (e.g.,diisopropyl azodicarboxylate (DIAD), etc.) in a solvent inert to thereaction (e.g., THF, etc.).

Step I-6 is a step of obtaining Intermediate I-5 from Compound 6i. Thisstep can be performed by reacting Compound 6i with a reducing agent(e.g., triphenylphosphine, etc.), in a solvent inert to the reaction(e.g., THF, etc.), and then treating the resultant with water, andheating them.

Next, the production methods of Intermediates II and II′ will bedescribed. The production methods shown below are merely examples andshould not be construed as limited to these.

Intermediates II and II′ are known, or are produced using a knowncompound as a starting material according to a known method or a methodanalogous thereto. Examples of known document include Nat. Chem. Biol.2012, 8, 277-284., Cancer cell. 2015, 27, 589-602., J. Med. Chem. 2016,59(3), 892-913., WO 2017//214367, WO 2016/195776, WO 2012/097013, J.Heterocyclic Chem. 2005, 42(4), 509-513., J. Med. Chem. 2001, 44(17),2695-2700. and the like.

When Intermediate II is represented by the following Compound II-1 orII′-1, it can also be produced according to the following method (MethodJ). The raw material, Compound 1j is known, or is produced using a knowncompound as a starting material according to a known method or a methodanalogous thereto. Examples of known document include WO 2004/007491.

Method J

In the scheme, R^(j1) is a chlorine atom or a methoxy group. R^(j2) andR^(j3) are both carbon atoms, or R^(j2) is a sulfur atom and R^(j3) is abond.

Step J-1 is a step of obtaining Compound 2j from Compound 1j. This stepcan be performed by reacting Compound 1j with(trifluoromethyl)trimethylsilane (Ruppert reagent) and a reagent to be afluoride ion source (e.g., tetrabutylammonium fluoride, etc.), in asolvent inert to the reaction (e.g., THF, etc.).

Step J-2 is a step of obtaining Compound 3j from Compound 2j. This stepcan be performed by treating Compound 2j with an acid (e.g.,hydrochloric acid, etc.) in a solvent inert to the reaction (e.g.,tetrahydrofuran, etc.).

Step J-3 is a step of obtaining Compound 4j from Compound 3j. This stepcan be performed by reacting Compound 3j with phenylchlorothionocarbonate and a base (e.g., TEA, etc.) in a solvent inert tothe reaction (e.g., dichloromethane, etc.).

Step J-4 is a step of obtaining Intermediate II-1 from Compound 4j. Thisstep can be performed by reacting Compound 4j with a radical reducingagent (e.g., tributyltin hydride, etc.) and a radical initiator (e.g.,azobis(isobutyronitrile), etc.) in a solvent inert to the reaction(e.g., toluene, etc.).

Step J-5 is a step of obtaining Intermediate II′-1 wherein R^(j1) is ahydroxy group from Intermediate II-1 wherein R^(j1) is a methoxy group.This step can be performed by treating Intermediate II-1 wherein R^(j1)is a methoxy group with an acid (e.g., hydrochloric acid, etc.) in asolvent inert to the reaction (e.g., THF, etc.).

When Intermediate III is represented by the following Compound III-1 orIII-2, it can also be produced according to the following method (MethodK). The raw material, Compound 1k is known, or is produced using a knowncompound as a starting material according to a known method or a methodanalogous thereto. Examples of known document include Eur. J. Org. Chem.2013, 17, 3477-3493.

Method K

Steps K-1 to K-5 can be performed in the same manner as in Steps F-1 toF-5, and Step K-6 can be performed in the same manner as in Step A-1. Inthe purification process of Step K-6, various isomers by-produced in aseries of steps can also be removed by using an optically active columnsuch as CHIRALPAK (registered trademark, Daicel Co., Ltd.)-IA, IB, IC,ID and the like. As the developing solvent, n-hexane, ethanol, isopropylalcohol and the like can be used.

Step K-7 is a step of obtaining Intermediates III-1 and III-2 fromCompound 7k. This step can be performed by reacting Compound 7k with acatalyst (e.g., osmium tetraoxide, etc.) and an oxidizing agent (e.g.,4-methylmorpholine N-oxide, etc.) in a solvent inert to the reaction(e.g., a mixed solvent of acetone and water, etc.).

When Intermediate III is represented by the following Compound III-3, itcan also be produced according to the following method (Method L). Theraw material, Compound 11 is known, or is produced using a knowncompound as a starting material according to a known method or a methodanalogous thereto. Examples of known document include Bioorg. Med. Chem.2006, 14, 2242-2252.

Method L

Steps L-1 to Step L-4 can be performed in the same manner as in StepsI-3 to I-6, and Step L-5 can be performed in the same manner as in StepA-1, and Step L-6 can be performed in the same manner as in Step G-2.

Step L-7 is a step of obtaining Compound 51 from Compound 41. This stepcan be performed by reacting Compound 41 with Compound 71 in thepresence of a condensing agent (e.g., COMU, etc.) and a base (e.g.,DIPEA, etc.), in a solvent inert to the reaction (e.g., DMF, etc.).

Step L-8 is a step of obtaining Compound 61 from Compound 51. This stepcan be performed by treating Compound 51 with an acid (e.g.,trifluoroacetic acid, etc.), in a solvent inert to the reaction (e.g.,dichloromethane, etc.).

Step L-9 is a step of obtaining Intermediate III-3 from Compound 61.This step can be performed by reacting Compound 61 with a phosphinecompound (e.g., triphenylphosphine, etc.), hexachloroethane and a base(e.g., triethylamine, etc.) in a solvent inert to the reaction (e.g.,dichloromethane, etc.).

When Intermediate III is represented by the following Compound III-4, itcan be produced according to the following method (Method M). Compound1m can be synthesized according to a known method (WO 2017//214367). Theboronic acid (3m) is a known compound, or can be produced using a knowncompound as a starting material, according to a known method or a methodanalogous thereto.

Method M

In the scheme, R^(n1) is an aromatic ring group.

Step M-1 is a step of obtaining Compound 2m from Intermediate I andCompound 1m. This step can be performed in the same manner as in StepA-1.

Step M-2 is a step of obtaining Intermediate III-4 from Compound 2m.This step can be performed by heating Compound 2m using a metal catalyst(e.g., tetrakis(triphenylphosphine)palladium, etc.) and a base (e.g.,sodium carbonate, etc.), under inert gas atmosphere, in a solvent inertto the reaction (e.g., a mixed solvent of dioxane and water, etc.). Inthis step, a boronate may be used instead of the boronic acid (3m).

When Intermediate III is represented by the following Compound III-5, itcan be produced, for example, according to the following method (MethodN). Compound in can be produced in the same manner as in Step A-1 ofMethod A.

Method N

In the scheme, R^(n1) is a C₁₋₆ alkyl group, a p-methoxybenzyl group orthe like.

Step N is a step of obtaining Intermediate III-5 from Compound in. Thisstep can be performed by reacting Compound 25 In with an amine (e.g.,methylamine, para-methoxybenzylamine, etc.) under heating (desirably,heating above the boiling point of the solvent using a microwave reactoretc.) in a solvent inert to the reaction (e.g., butyronitrile, etc.).

Next, the production methods of Intermediate V will be described.Intermediate V is known, or is produced using a known compound as astarting material according to a known method or a method analogousthereto. Examples of known document include Cancer cell. 2015, 27,589-602, J. Med. Chem. 2016, 59(3), 892-913, WO 2007/118041, WO2014/164749 and the like.

Since Intermediate V has a functional group represented by R¹C(═O)— (aformyl group or a C₁₋₆ alkylcarbonyl group) on Ring Q¹, it can also bederived from a precursor having a group that can be easily converted tothe functional group (in the case of a formyl group, examples thereofinclude a hydroxymethyl group, a C₁₋₆ alkoxy carbonyl group, a carboxygroup and the like, and in the case of a C1-6 alkylcarbonyl group,examples thereof include an acetyl group, an ethanoyl group and thelike).

Intermediate V can also be produced according to Method O to Method Y.

Intermediate V is represented by the following Compound V-1, it can alsobe produced, for example, according to Method O.

Method O

In the scheme, R^(o1) is a halogen atom (e.g., chlorine, bromine, oriodine) or a trifluoromethylsulfonyloxy group. R^(o2) is a functionalgroup that can be easily converted to a formyl group, and examplesthereof include a hydroxylmethyl group, a C₁₋₆ alkoxy carbonyl group, acarboxy group, an acetal group and the like. Ring Q⁴ is a heterocyclecontaining a nitrogen atom in the ring (the heterocycle optionally hassubstituent(s)), and examples thereof include a piperidine ring and thelike.

Step O-1 is a step of obtaining Compound 3o from Compounds 1o and 2o.This step can be performed by heating Compounds 1o and 2o under inertgas atmosphere in the presence of a metal catalyst (e.g., a combinationof tris(dibenzylideneacetone)dipalladium(0) and(±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl, etc.) and a base (e.g.,sodium tert-butoxide, cesium carbonate, triethylamine, etc.), in asolvent inert to the reaction (e.g., toluene, etc.).

Step O-2 is a step of converting Compound 3o to Intermediate V-1. Forexample, when R^(o2) is a hydroxymethyl group, it can be performed byreacting Compound 3o with an oxidizing agent (pyridinium chlorochromate,Dess-Martin periodinane, manganese(IV) oxide, etc.) in a solvent inertto the reaction (e.g., dichloromethane, chloroform, DMSO, etc.). WhenR^(o2) is the other functional group (e.g., a C₁₋₆ alkoxy carbonylgroup, a carboxy group, an acetal group, etc.), the conversion of thefunctional group to a formyl group can be performed according to themethods described in the above-mentioned “ORGANIC FUNCTIONAL GROUPPREPARATIONS”, “Comprehensive Organic Transformations” etc., and thelike.

When Intermediate V is represented by the following Compound V-2, it canalso be produced, for example, according to Method P.

Method P

In the scheme, R^(p1) is a halogen atom (e.g., chlorine, bromine, oriodine) or a trifluoromethylsulfonyloxy group. R^(p2) is a functionalgroup that can be easily converted to a formyl group, and examplesthereof include a hydroxylmethyl group, a C₁₋₆ alkoxy carbonyl group, acarboxy group, an acetal group and the like. Ring Q⁵ is a heterocyclecontaining a nitrogen atom in the ring (the heterocycle optionally hassubstituent(s)), and examples thereof include a dihydroindole ring andthe like.

Step P-1 is a step of obtaining Compound 3p from Compound 1p andCompound 2p. This step can be performed in the same manner as in StepO-1.

Step P-2 is a step of obtaining Intermediate V-2 from Compound 3p. Forexample, when R² is an acetal group, it can be performed by treatingCompound 3p with an acid (e.g., hydrochloric acid, etc.) in a solventinert to the reaction (e.g., THF, etc.).

Intermediate V is represented by the following Compound V-3, it can alsobe produced, for example, according to Method Q.

Method Q

In the scheme, one of R^(q1) and R^(q2) is an amino group, and the otherof R^(q1) and R^(q2) is a halogen atom (e.g., chlorine, bromine, oriodine) or a trifluoromethylsulfonyloxy group. R^(q3) is a formyl groupor a functional group that can be easily converted to a formyl group,and examples thereof include a hydroxylmethyl group, a C₁₋₆ alkoxycarbonyl group, a carboxy group, an acetal group and the like.

Step Q-1 is a step of obtaining Compound 3q from Compound 1q andCompound 2q. This step can be performed by heating Compound 1q andCompound 2q under inert gas atmosphere in the presence of a metalcatalyst (e.g., a combination oftris(dibenzylideneacetone)dipalladium(0) and tert-butylphosphine, ortetrakis(triphenylphosphine)palladium(0), etc.) and a base (e.g., sodiumtert-butoxide, sodium carbonate, etc.), in a solvent inert to thereaction (e.g., a mixed solvent of toluene, dioxane and water, a mixedsolvent of dimethoxyethane and water, etc.).

Step Q-2 is a step of obtaining Intermediate V-3 from Compound 3q. Forexample, when R^(q3) is an acetal group, it can be performed by treatingCompound 3q with an acid (e.g., hydrochloric acid, etc.) in a solventinert to the reaction (e.g., THF, etc.). When R^(q3) is a formyl group,this step is not required.

Intermediate V is represented by the following Intermediate V-4, it canalso be produced, for example, according to Method R.

Method R

In the scheme, one of R^(r1) and R^(r2) is a halogen atom (e.g.,chlorine, bromine, or iodine) or a trifluoromethylsulfonyloxy group, andthe other of R^(r1) and R^(r2) is a borono group, a dialkoxyboranylgroup (e.g., a dimethoxyboranyl group, etc.), a dioxaborolanyl group (a4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, etc.) and the like.R^(r3) is a formyl group or a functional group that can be easilyconverted to a formyl group, and examples thereof include ahydroxylmethyl group, a C₁₋₆ alkoxy carbonyl group, a carboxy group, anacetal group and the like.

Step R-1 is a step of obtaining Compound 3r from Compound 1r andCompound 2r. This step can be performed by heating Compound 1r andCompound 2r under inert gas atmosphere in the presence of a metalcatalyst (e.g., a combination oftris(dibenzylideneacetone)dipalladium(0) and tert-butylphosphine, ortetrakis(triphenylphosphine)palladium(0), etc.) and a base (e.g., sodiumtert-butoxide, sodium carbonate, tripotassium phosphate, etc.), in asolvent inert to the reaction (e.g., a mixed solvent of toluene, dioxaneand water, a mixed solvent of dimethoxyethane and water, etc.).

Step R-2 is a step of obtaining Intermediate V-4 from Compound 3r. Forexample, when R³ is a C₁₋₆ alkoxy carbonyl group, this step can beperformed according to the below Method V. When R³ is a formyl group,this step is not required.

The raw material compounds used in Method O to Method R are known, orare produced using a known compound as a starting material according toa known method or a method analogous thereto. Examples of known documentinclude WO 2014/078813, Synlett. 2015, 26(7), 953-959, J. Med. Chem.2014, 57(19), 8086-8098, Eur. J. Inorg. Chem. 2015, 28, 4666-4677, WO2004/108690, WO 2015/0291572, WO 2010/141796, WO 2013/093849, J. Med.Chem. 2014, 57(19), 8086-8098, J. Org. Chem. 2014, 79, 10311-10322, WO2011/109267, WO 2007/013673, Tetrahedron. 2015, 71(49), 9240-9244,“ORGANIC FUNCTIONAL GROUP PREPARATIONS”, the 2nd edition, ACADEMICPRESS, INC., 1989, “Comprehensive Organic Transformations”, VCHPublishers Inc., 1989 and the like.

Compound 1r can also be produced according to the following Method S orMethod T.

When Compound 1r is represented by the following Compound 2s or 3s, itcan be produced, for example, according to Method S.

Method S

In the scheme, R^(s1), R^(s2) and R^(s3) are each independently a C₁₋₆alkyl group (e.g., a methyl group, an ethyl group, etc.), an aromaticring group (e.g., pyridyl group, etc.) optionally containingheteroatom(s), or a saturated heterocyclic group (e.g., piperidinylgroup, etc.) and the like, or R^(s2) and R^(s3) are optionally takentogether with the nitrogen atom to which R^(s2) and R^(s3) are bonded toform a ring.

Step S-1 is a step of obtaining Compound 2s from Compound 1s. This stepcan be performed by reacting Compound is with Compound 4s in thepresence of a base (e.g., sodium hydride, potassium carbonate, etc.), ina solvent inert to the reaction (e.g., toluene, DMF, etc.).

Step S-2 is a step of obtaining Compound 3s from Compound 1s. This stepcan be performed by reacting Compound is with Compound 5s in thepresence of a base (e.g., diisopropylamine, etc.) in a solvent inert tothe reaction (e.g., THF, etc.).

When Compound 1r is represented by any of the following Compounds 3t to5t, it can also be produced, for example, according to Method T.

Step T-1 is a step of obtaining Compound it and Compound 2t fromCompound 1s. This step can be performed by reacting Compound is with abase (e.g., potassium acetate, etc.) in a solvent inert to the reaction(e.g., a mixed solvent of acetic acid and water, etc.), and then heatingthe mixture. The resulting Regioisomers 1t and 2t can be separated fromeach other, for example, by utilizing the difference in solubility.

Step T-2 is a step of obtaining Compound 3t from Compound it. This stepcan be performed by reacting Compound it with a base (e.g., aqueouspotassium hydroxide solution, etc.) and a difluoromethylating agent(e.g., difluoromethyltrifluoromethanesulfonic acid, etc.), in a solventinert to the reaction (e.g., acetonitrile, etc.).

Step T-3 is a step of obtaining Compounds 4t and 5t from Compound 2t.This step can be performed in the same manner as in Step T-2. Theresulting Isomers 4t and 5t can be separated from each other, forexample, by a method using silica gel column chromatography and thelike.

When Intermediate V is represented by the following Compound V-5, it canbe produced, for example, according to the following method (Method U).

Method U

In the scheme, Ring Q⁶ is a heterocycle containing a nitrogen atom inthe ring (the heterocycle optionally has substituent(s)), and examplesthereof include a piperidine ring, an azepane ring and the like.

Step U is a step of obtaining Intermediate V-5 from Compound 1u andCompound 2u. This step can be performed by reacting Compound 1u andCompound 2u with an amine (e.g., DIPEA, etc.) in a solvent inert to thereaction (e.g., dichloromethane, etc.).

When Intermediate V is represented by the following Compound V-6, it canalso be produced, for example, according to the following method (MethodV). The starting material is known, or is produced using a knowncompound as a starting material according to a known method or a methodanalogous thereto. Examples of known document include WO 2014/114186, WO2005/108399, J. Med. Chem. 2016, 59(18), 8233-8262, WO 2005/032488,Science. 2016, 352(6291), 1304-1308, Bioorg. Med. Chem. Let. 2006,16(19), 4987-4993, Helvetica Chimica Acta. 2007, 90(6), 1043-1068,Tetrahedron. 2015, 71(49), 9240-9244, “ORGANIC FUNCTIONAL GROUPPREPARATIONS”, the 2nd edition, ACADEMIC PRESS, INC., 1989,“Comprehensive Organic Transformations”, VCH Publishers Inc., 1989, andthe like.

Method V

In the scheme, R^(v1) is a hydrogen atom or an alkyl group (e.g., amethyl group, an ethyl group, etc.).

Step V-1 is a step of obtaining Compound 2v from Compound 1v. This stepcan be performed, for example, by reacting Compound 1v with a reducingagent (e.g., lithium aluminium hydride, lithium borohydride, etc.) in asolvent inert to the reaction (e.g., THF, etc.).

Step V-2 is a step of obtaining Intermediate V-6 from Compound 2v. Thisstep can be performed, for example, by reacting Compound 2v with anoxidizing agent (e.g., pyridinium chlorochromate, Dess-Martinperiodinane, manganese(IV) oxide, etc.) in a solvent inert to thereaction (e.g., dichloromethane, chloroform, DMSO, etc.).

Step V-3 is a step of obtaining Intermediate V-6 from Compound 1v. Thisstep can be performed by reacting Compound 1v with a reducing agent(e.g., diisobutylaluminium hydride, etc.) in a solvent inert to thereaction (e.g., dichloromethane, etc.).

When Intermediate V is represented by the following Compound V-7, it canbe produced, for example, according to Method W.

Method W

In the scheme, R^(w1) is a C₁₋₆ alkyl group optionally havingsubstituent(s) (e.g., a 3-(tert-butoxycarbonylamino)propyl group, a1-methylpyrazol-4-ylmethyl group, etc.).

Step W is a step of obtaining Intermediate V-7 from Compounds 1w and 2w.This step can be performed in the same manner as in Step I-1.

When Intermediate V has a functional group, Intermediate V can also beused after converting the functional group to a desired functional groupby a known method. Examples of known method include the methodsdescribed in “ORGANIC FUNCTIONAL GROUP PREPARATIONS”, “ComprehensiveOrganic Transformations” and “Protective Groups in Organic Synthesis(the 5th edition, 2014)”, and the methods described in J. Med. Chem.2014, 57(18), 7590-7599, Synlett. 2015, 26(7), 953-959, WO 2013/013503,Angew. Chem. Int. Ed. 2015, 54(33), 9668-9672, Org. Lett. 2012, 14(14),3700-3703, WO 2017//100668 and the like, and various reactions such asreduction of a nitro group, acroylation, sulfonylation, methylation,deprotection and the like can be performed.

When the compound represented by the general formula (1) has afunctional group, the functional group can also be converted to adesired functional group by a known method. For example, protection ordeprotection, conversion or modification of functional groups, and thelike can be performed. Examples of known method include the methodsdescribed in “ORGANIC FUNCTIONAL GROUP PREPARATIONS”, “ComprehensiveOrganic Transformations” and “Protective Groups in Organic Synthesis(the 5th edition, 2014)”, and the methods described in J. Med. Chem.2014, 57(18), 7590-7599, Angew. Chem. Int.Ed.2017, 56(21), 5886-5889 andthe like.

The compound produced by the above-mentioned method can be isolated andpurified by a known method, for example, extraction, precipitation,distillation, chromatography, fractional recrystallization,recrystallization and the like.

The inhibitory activity of the cell-free binding between menin and MLLcan be measured by employing the chemiluminescent AlphaLISA (registeredtrademark, PerkinElmer) method described in the following ExperimentalExample 1. Alternatively, as general methods for evaluatingprotein-protein interaction, time-resolved fluorescence resonance energytransfer (TR-FRET) method in which fluorescence energy transfer from adonor-binding protein having long-lived fluorescence to anacceptor-binding protein is detected by FRET (Fluorescence ResonanceEnergy Transfer), surface plasmon resonance (SPR) method, and the likecan be employed.

The cell growth inhibitory activity of the compound of the presentinvention or a pharmaceutically acceptable salt thereof can be examinedby employing a growth inhibitory test method conventionally employed bythose skilled in the art. The cell growth inhibitory activity can beexamined, for example, as described in the following ExperimentalExample 2, by comparing the degree of cell growth obtained in thepresence of a test compound with that obtained in the absence of thetest compound. The degree of growth can be examined, for example, byusing a test system for measuring living cells. Examples of method formeasuring living cells include a [³H]-thymidine uptake assay, a BrdUmethod and an MTT assay.

The antitumor activity in vivo can be examined by an antitumor testmethod conventionally employed by those skilled in the art. For example,as described in the following Experimental Examples 3-1 to 3-4, varioustumor cells are transplanted into a mouse, a rat or the like, and afterconfirming engraftment of the transplanted cells, the compound of thepresent invention is orally or intravenously administered. After severaldays or several weeks, the tumor growth in the non-administration groupand that in the compound administration group are compared, so that theantitumor activity in vivo according to the present invention can beconfirmed.

The compound of the present invention or a pharmacologically acceptablesalt thereof can be used together with other antitumor agents. Examplesthereof include alkylating agents, antimetabolites, antitumorantibiotics, antitumor plant components, BRMs (biological responsemodifiers), hormones, vitamins, antitumor antibodies, molecular targetdrugs, other antitumor agents and the like.

More specifically, examples of the alkylating agent include alkylatingagents such as nitrogen mustard, nitrogen mustard N-oxide, chlorambuciletc.; aziridine-based alkylating agents such as carboquone, thiotepa,etc.; epoxide-based alkylating agents such as dibromomannitol, dibromodulcitol etc.; nitrosourea-based alkylating agents such as carmustine,lomustine, semustine, nimustine hydrochloride, streptozocin,chlorozotocin, ranimustine etc.; and busulfan, improsulfan tosylate,dacarbazine and the like.

Examples of the antimetabolite include purine antimetabolites such as6-mercaptopurine, 6-thioguanine, thioinosine, etc.; pyrimidineantimetabolites such as fluorouracil, tegafur, tegafur-uracil, carmofur,doxifluridine, broxuridine, cytarabine, enocitabine etc.; folic acidantimetabolites such as methotrexate, trimetrexate etc.; and the like.

Examples of the antitumor antibiotic include mitomycin C, bleomycin,peplomycin, daunorubicin, aclarbicin, doxorubicin, idarubicin,pirarubicin, THP-adriamycin, 4′-epidoxorubicin or epirubicin,chromomycin A3, and actinomycin D and the like.

Examples of the antitumor plant component include vinca alkaloids suchas videsine, vincristine, vinblastine etc.; taxanes such as paclitaxel,docetaxel etc.; and epipodophyllotoxins such as etoposide, teniposideetc.

Examples of the BRM include tumor necrosis factor, indomethacin and thelike.

Examples of the hormone include hydrocortisone, dexamethasone,methylprednisolone, prednisolone, prasterone, betamethasone,triamcinolone, oxymetholone, nandrolone, methenolone, fosfestrol,ethinylestradiol, chlormadinone, mepitiostane, medroxyprogesterone andthe like

Examples of the vitamin include vitamin C, vitamin A and the like.

Examples of the antitumor antibody and molecule target drug includevenetoclax, trastuzumab, rituximab, cetuximab, nimotuzumab, denosumab,bevacizumab, infliximab, ipilimumab, nivolumab, pembrolizumab, avelumab,pidilizumab, atezolizumab, ramucirumab, imatinib mesylate, dasatinib,gefitinib, erlotinib, osimertinib, sunitinib, lapatinib, dabrafenib,trametinib, cobimetinib, pazopanib, palbociclib, panobinostat,sorafenib, crizotinib, vemurafenib, quizartinib, bortezomib,carfilzomib, ixazomib, midostaurin, gilteritinib and the like.

Examples of the other antitumor agent include cisplatin, carboplatin,oxaliplatin, tamoxifen, letrozole, anastrozole, exemestane, toremifenecitrate, fulvestrant, bicalutamide, flutamide, mitotane, leuprorelin,goserelin acetate, camptothecin, ifosfamide, cyclophosphamide,melphalan, L-asparaginase, aceglatone, schizophyllan, picibanil,procarbazine, pipobroman, neocarzinostatin, hydroxyurea, ubenimex,thalidomide, lenalidomide, pomalidomide, eribulin, tretinoin, krestinand the like.

The pharmaceutical composition of the present invention comprises thecompound of the present invention or a pharmaceutically acceptable salt,and a pharmaceutically acceptable carrier, and can be administered asvarious injections such as intravenous injection, intramuscularinjection, subcutaneous injection etc., or by various methods such asoral administration, transdermal administration etc. Thepharmaceutically acceptable carrier means a pharmaceutically acceptablematerial (e.g., an excipient, a diluent, an additive, a solvent, etc.),which is involved in transporting the compound of the present inventionor the composition containing the compound of the present invention fromone organ to another organ.

A formulation containing the compound of the present invention or apharmacologically acceptable salt thereof as an active ingredient isprepared by using an additive used in a conventional formulation, suchas a carrier, an excipient etc. Administration of the compound of thepresent invention can be oral administration in the form of a tablet, apill, a capsule, a granule, a powder, a liquid or the like, orparenteral administration in the form of an injection (such asintravenous injection, intramuscular injection etc.), a suppository, atransdermal agent, a nasal agent, an inhalant or the like. The dose andthe number of doses of the compound of the present invention areappropriately determined depending on individual cases in considerationof the symptoms, and the age, the sex or the like of an administrationtarget. The dose is usually 0.001 mg/kg to 100 mg/kg per dose for oraladministration to an adult, and usually 0.0001 mg/kg to 10 mg/kg perdose for intravenous administration to an adult. The number of doses isusually once to six times per day, or once per day to once per 7 days.

A solid formulation for oral administration of the present invention canbe a tablet, a powder, a granule or the like. Such a formulation can beproduced by mixing one or more active substances with an inertexcipient, lubricant, disintegrant, dissolution assisting agent and thelike, according to a conventional method. The excipient can be, forexample, lactose, mannitol or glucose. The lubricant can be, forexample, magnesium stearate. The disintegrant can be, for example,sodium carboxymethyl starch. A tablet or pill can be coated with a sugarcoating or a gastric-soluble or enteric coating agent if necessary.

A liquid formulation for oral administration can be a pharmaceuticallyacceptable emulsion, liquid, suspension, syrup, elixir and the like.Such a formulation contains a generally used inert solvent (e.g.,purified water, ethanol), and may further contain a solubilizing agent,a wetting agent, a suspending agent, a sweetener, a flavoring agent, anaromatic or a preservative.

An injection for parenteral administration can be an aseptic aqueous ornon-aqueous liquid, suspension or emulsion. An aqueous solvent forinjection can be, for example, distilled water or a normal salinesolution. A non-aqueous solvent for injection can be, for example,propylene glycol, polyethylene glycol, vegetable oils such as olive oil,alcohols such as ethanol, or polysorbate 80 (pharmacopoeia name). Such aformulation may further contain a tonicity agent, a preservative, awetting agent, an emulsifier, a dispersant, a stabilizer or adissolution assisting agent. Such a formulation can be sterilized, forexample, by filtration through a bacteria retention filter, blendingwith a bactericide, or radiation exposure. Alternatively, a compositionobtained by dissolving or suspending an aseptic solid composition inaseptic water or an injection solvent before use can also be used as aformulation.

EXAMPLE

Hereinafter, the present invention will be described in more detail withreference to Reference Examples and Examples, but the scope of thepresent invention is not limited to these examples, and these examplesare not restrictively interpreted in any sense. In addition, thereagents, solvents and starting materials used herein are readilyavailable from commercially available sources, unless otherwisespecified.

The proton nuclear magnetic resonance spectrum (¹H-NMR) was measuredusing 400 MHz nuclear magnetic resonance spectrometer manufactured byJEOL, or 400 MHz nuclear magnetic resonance spectrometer manufactured byVarian. The spectral data indicates significant peaks, showing thechemical shifts (which are shown as relative ppm (δ) from atetramethylsilane peak), the number of protons, and the multiplicity ofpeak splitting (which are shown as s: singlet; d: doublet; t: triplet;q: quartet; quint: quintet; m: multiplet; br: broad; br s: broadsinglet, etc.), and further, if specified, the coupling constant J value(unit: Hz).

The mass spectrum (MS m/z) was measured using an electrospray ionizationmethod (ESI) or an atmospheric pressure chemical ionization method(APCI). The mass spectral data was shown regarding the maximumionization peak (corresponding to the maximum UV absorption peak inalmost all cases) obtained after passing through a reverse phase highperformance liquid chromatography column (Agilent System; column:Develosil Combi-RP-5, 2.0×50 mm, Cadenza CD-C18, 3.0×75 mm, or ZORBAXSB-C18, 1.8 μm, 2.1×50 mm; solvent: 0.1% formic acid-containingacetonitrile/water system, or 0.01% trifluoroacetic acid-containingacetonitrile/water system).

The silica gel column chromatography was performed by using acommercially available packed column and an automatic preparativepurification system (e.g., SP1 manufactured by Biotage, EPCLC-W-Prep2XYmanufactured by Yamazen, Purif-α2 manufactured by Shoko Science, etc.),and multiple types of solvents used for the mobile phase were merelydescribed. The elution is performed under observation by thin layerchromatography (TLC). As a TLC plate, silica gel 60 F₂₅₄ or 60 NH₂ F₂₅₄smanufactured by Merck, NH₂ silica gel 60 F₂₅₄ plate manufactured by WakoPure Chemical Industries, Ltd. or CHROMATOREX NH TLC manufactured byFuji Silysia Chemical Ltd. was used. As a developing solvent, the mobilephase used in the column chromatography was used. As a detection method,a UV detector or a color-developing reagent was employed. In ReferenceExamples and Examples, the “amino silica gel” refers to silica gel whosesurface is chemically modified by a functional group having an aminogroup (e.g., Purif-Pack (registered trademark, Shoko Scientific)-EX, NHseries etc.).

The preparative thin layer chromatography (PTLC) was performed by usingsilica gel 60 F₂₅₄ plate manufactured by Merck, or silica gel 70 PF₂₅₄plate or NH₂ silica gel 60 F₂₅₄ plate manufactured by Wako Pure ChemicalIndustries, Ltd., and multiple types of solvents used for the mobilephase were merely described.

The preparative high-performance liquid chromatography (preparativeHPLC) was performed by using reverse-phase column (Develosil Combi-RP-5)manufactured by Nomura Chemical, and 0.1% formic acid-containingacetonitrile/water was used for the mobile phase.

The solvent amount and solvent ratio, the conversion timing and thegradient method used in these chromatographies are not described herein.However, the purification and/or separation method used herein can bereproduced by ordinary knowledge and/or techniques of chemicalsynthesis.

In Examples, the equipment and measurement conditions in the powderX-RAY diffraction measurement are as follows:

Model: Rigaku Rint TTR-III

Sample holder: non-reflecting sample holderSample amount: appropriate amountX-RAY generation conditions: 50 kV, 300 mAWavelength: 1.54 Å (Cu-Kα rays)Measurement temperature: room temperatureScanning speed: 20°/minScanning range: 2 to 40°Sampling width: 0.02°Analytical procedure: Several mg of the test compound was taken with aspatula, placed on a non-reflecting sample holder, and flattened with amedicine wrapping paper. Thereafter, the peak pattern was analyzed underthe above conditions.

In the following Reference Examples and Examples, the racemates,optically active substances, geometric isomers, and steric notations aredescribed according to the following criteria for convenience. (1) Thesteric notation in the structural formula is shown using a wedge-shapedline (bond forwards out of the plain of the page), a broken line (bondbackwards out of the plain of the page), a solid line (bond in the plainof the page) or a wavy line (bond not specifying configuration). (2)When “racemate” is described together with the structural formula of thecompound, it indicates that the compound represented by the structuralformula is a racemate (an equal amount mixture with an enantiomer). When“diastereomeric mixture” is described together with the structuralformula of the compound, it indicates that the compound represented bythe structural formula is a mixture of diastereomers (derived from theconfiguration of bonds indicated by the wavy line). When “cis-transmixture” is described together with the structural formula of thecompound, it indicates that the compound represented by the structuralformula is a mixture of geometric isomers. In addition, unless otherwisespecified in the structural formula and compound name, it indicates thatthe compound represented by the structural formula is a singlestereoisomer (including an optically active substance). (3) When“(racemic)” is attached after the compound name, it indicates that thecompound indicated by the compound name is a racemic (an equal amountmixture with an enantiomer). When “(diastereomeric mixture)” is attachedafter the compound name, it indicates that the compound indicated by thecompound name is a mixture of diastereomers (derived from theconfiguration of the bond indicated by the wavy line). When “(cis-transmixture)” is attached after the compound name, it indicates that thecompound indicated by the compound name is a mixture of geometricisomers.

In the following Reference Examples and Examples, the notations of “HCl”described together with the structural formula of the compound and“hydrochloric acid” attached the compound name indicate that thecompound represented by the structural formula or the compound indicatedby the compound name and hydrogen chloride can form the hydrochloride invarious ratios, not necessarily in a 1:1 ratio (for example, includingmonohydrochloride, dihydrochloride, trihydrochloride, etc.).

Reference Example A-1 benzyl[(1R,3R,4S)-3-hydroxy-4-(methylamino)cyclopentyl]carbamate Step 1 methyl(1R,4S)-4-aminocyclopent-2-ene-1-carboxylate hydrochloride

To a mixture of (1S)-(+)-2-azabicyclo[2.2.1]hept-5-en-3-one (CAS:130931-83-8) (98.6 g) and methanol (300 mL) was added thionyl chloride(CAS: 7719-09-7) (40 mL) over 50 min at 0° C., and the mixture wasstirred at 0° C. for 2 hr. The reaction mixture was concentrated underreduced pressure, and the obtained solid was suspended in ethyl acetate,and collected by filtration to give the title compound (158 g) as asolid. ¹H-NMR (DMSO-D₆) δ: 1.88-1.99 (1H, m), 2.48-2.62 (1H, m),3.64-3.74 (4H, m), 4.15-4.23 (1H, m), 5.84-5.90 (1H, m), 6.07-6.11 (1H,m), 8.20 (3H, br s).

Step 2 methyl(1R,4S)-4-[(tert-butoxycarbonyl)amino]cyclopent-2-ene-1-carboxylate

To a mixture of the compound (158 g) obtained in the above Step 1, THF(700 mL) and water (280 mL) were added di-tert-butyl dicarbonate (CAS:24424-99-5) (194 g) and sodium carbonate (CAS: 497-19-8) (104 g) at 0°C., and the mixture was stirred overnight at room temperature. Water wasadded to the reaction solution, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated brine, dried overanhydrous sodium sulfate, and concentrated under reduced pressure togive the title compound (214 g) as an oil. ¹H-NMR (CDCl₃) δ: 1.44 (9H,s), 1.82-1.91 (1H, m), 2.46-2.57 (1H, m), 3.45-3.52 (1H, m), 3.71 (3H,s), 4.75-4.84 (1H, m), 4.86-4.96 (1H, m), 5.84-5.91 (2H, m).

MS (m/z): 142 (M-Boc+H)⁺.

Step 3 methyl(3aS,5S,6S,6aS)-6-bromo-2-oxohexahydro-2H-cyclopenta[d][1,3]oxazole-5-carboxylate

To a mixture of the compound (214 g) obtained in the above Step 2, THF(700 mL) and water (70 mL) was added N-bromosuccinimide (CAS: 128-08-5)(174 g) at 0° C., and the mixture was stirred overnight at roomtemperature. The reaction solution was concentrated under reducedpressure, and ethyl acetate was added thereto. The organic layer waswashed with 1N hydrochloric acid, saturated aqueous sodiumhydrogencarbonate solution, saturated aqueous sodium thiosulfatesolution and saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was suspended in ethylacetate/n-hexane, and the solid was collected by filtration. Theobtained solid was again suspended in ethyl acetate/n-hexane, andcollected by filtration to give the title compound (113 g) as a solid.The filtrate was concentrated under reduced pressure, the residue wassuspended in ethyl acetate/n-hexane, and the solid was collected byfiltration. The obtained solid was again suspended in ethylacetate/n-hexane, and collected by filtration. Ethyl acetate/n-hexaneand water were added to the obtained solid, and the mixture wassubjected to liquid separation. The organic layer was dried overanhydrous sodium sulfate, and concentrated under reduced pressure togive the title compound (11.0 g) as a solid. In total, the titlecompound (124 g) was obtained as a solid.

¹H-NMR (CDCl₃) δ: 2.39-2.47 (1H, m), 2.48-2.55 (1H, m), 3.21-3.25 (1H,m), 3.76 (3H, s), 4.42-4.45 (1H, m), 4.79 (1H, s), 5.16 (1H, dd, J=7.6,1.5 Hz), 5.95 (1H, br s).

MS (m/z): 264, 266 (M+H)⁺.

Step 4(3R,4S)-4-[(tert-butoxycarbonyl)amino]-3-hydroxycyclopent-1-ene-1-carboxylicacid

To a mixture of the compound (124 g) obtained in the above Step 3,methanol (580 mL) and water (580 mL) was added potassium hydroxide (CAS:1310-58-3) (106 g) at 0° C., and the mixture was stirred at 90° C. for18 hr. The reaction solution was concentrated under reduced pressure,THF (180 mL) and di-tert-butyl dicarbonate (CAS: 24424-99-5) (102 g)were added thereto at 0° C., and the mixture was stirred at roomtemperature for 4 hr. The mixture was neutralized with 5N hydrochloricacid and 2N hydrochloric acid at 0° C., and extracted five times withethyl acetate. The combined organic layers were dried over anhydroussodium sulfate, and the solvent was evaporated under reduced pressure.The obtained solid was suspended in ethyl acetate/n-hexane, collected byfiltration, and dried under reduced pressure to give the title compound.The filtrate was concentrated, and the obtained solid was suspended inethyl acetate/n-hexane, collected by filtration, and dried under reducedpressure to give the title compound. In total, the title compound (96.9g) was obtained as a solid.

¹H-NMR (DMSO-D₆) δ: 1.39 (9H, s), 2.30-2.45 (1H, m), 2.53-2.63 (1H, m),3.91-4.04 (1H, m), 4.47-4.54 (1H, m), 4.97-5.12 (1H, m), 6.35 (1H, d,J=7.9 Hz), 6.52 (1H, d, J=1.8 Hz), 12.53 (1H, br s).

MS (m/z): 242 (M−H)⁻.

Step 5(3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclopentane-1-carboxylicacid (diastereomer mixture)

A mixture of the compound (96.9 g) obtained in the above Step 4, 10%palladium on carbon wet (10.4 g) and methanol (780 mL) was stirred underhydrogen atmosphere for 7.5 hr. The reaction solution was filteredthrough Celite. The filtrate was concentrated under reduced pressure togive the title compound (105 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.38 (9H, s), 1.69-2.08 (4H, m), 2.59-2.72 (1H, m),3.52-3.73 (1H, m), 3.84-3.99 (1H, m), 6.15-6.29 (1H, m).

MS (m/z): 146 (M−Boc+H)⁺.

Step 6(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclopentane-1-carboxylicacid 2-methylpropan-2-amine salt

A mixture of the compound (105 g) obtained in the above Step 5,tert-butylamine (42.5 mL), methanol (90 mL) and tert-butyl methyl ether(675 mL) was allowed to stand in the refrigerator for 3 days. Theprecipitated solid was collected by filtration, and dried under reducedpressure to give the title compound (108 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.18 (9H, s), 1.35 (9H, s), 1.49-1.62 (1H, m),1.64-1.72 (1H, m), 1.72-1.85 (1H, m), 1.95-2.06 (1H, m), 2.44-2.53 (1H,m), 3.51-3.62 (1H, m), 3.71-3.79 (1H, m), 6.13 (1H, d, J=7.9 Hz).

MS (m/z) 146 (M−Boc+H)⁺.

Step 7(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclopentane-1-carboxylicacid

A mixture of the compound (108 g) obtained in the above Step 6 and water(200 mL) at 0° C. was adjusted to pH=3 with 2N hydrochloric acid (150mL). The mixture was extracted three times withdichloromethane/methanol=10/1, and the combined organic layers weredried over anhydrous sodium sulfate, and concentrated under reducedpressure to give the title compound (63.9 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.38 (9H, s), 1.70-1.84 (2H, m), 1.85-1.98 (1H, m),1.98-2.08 (1H, m), 2.61-2.71 (1H, m), 3.51-3.64 (1H, m), 3.84-3.92 (1H,m), 6.21 (1H, d, J=7.9 Hz).

MS (m/z): 146 (M-Boc+H)⁺.

Step 8 methoxymethyl(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylate

A mixture of the compound (61.6 g) obtained in the above Step 7,chloromethyl methyl ether (CAS: 107-30-2) (56.7 mL), DIPEA (CAS:7087-68-5) (262 mL), sodium iodide (75.2 g) and 1,2-dimethoxyethane(1000 mL) was heated under reflux for 1 hr. The reaction solution wasallowed to cool to room temperature, saturated brine and water wereadded thereto, and the mixture was extracted twice with ethyl acetate.The combined organic layers were dried over anhydrous sodium sulfate,and concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (66.5 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.87-1.98 (1H, m), 2.04-2.17 (1H, m),2.17-2.27 (1H, m), 2.27-2.38 (1H, m), 2.81-2.92 (1H, m), 3.37 (3H, s),3.46 (3H, s), 3.92-4.07 (2H, m), 4.61 (1H, d, J=6.7 Hz), 4.70 (1H, d,J=6.7 Hz), 5.09 (1H, d, J=10.0 Hz), 5.24 (2H, s).

MS (m/z): 234 (M-Boc+H)⁺.

Step 9(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylicacid

A mixture of the compound (66.5 g) obtained in the above Step 8, 1Naqueous sodium hydroxide solution (400 mL), methanol (20 mL) and THF(580 mL) was stirred at room temperature for 2 hr. 1N Hydrochloric acidwas added to the reaction solution at 0° C., and the mixture wasextracted twice with ethyl acetate. The combined organic layers werewashed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (53.6 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.86-1.99 (1H, m), 2.03-2.17 (1H, m),2.17-2.26 (1H, m), 2.26-2.38 (1H, m), 2.81-2.93 (1H, m), 3.38 (3H, s),3.91-4.08 (2H, m), 4.62 (1H, d, J=6.7 Hz), 4.70 (1H, d, J=6.7 Hz), 5.09(1H, d, J=6.7 Hz).

MS (m/z): 288 (M−H)⁻.

Step 10 benzyl tert-butyl[(1R,3S,4R)-4-(methoxymethoxy)cyclopentane-1,3-diyl]biscarbamate

A mixture of the compound (53.6 g) obtained in the above Step 9,diphenylphosphoryl azide (DPPA) (CAS: 26386-88-9) (51.9 mL), TEA (CAS:121-44-8) (33.4 mL) and toluene (730 mL) was stirred at 90° C. for 30min. Benzyl alcohol (38.3 mL) was added to the reaction solution, andthe mixture was stirred at 90° C. for 2.5 hr. The reaction solution wasallowed to cool to room temperature, water was added thereto, and themixture was extracted twice with ethyl acetate. The combined organiclayers were washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate) togive the title compound (63.2 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.35-1.50 (10H, m), 1.70-1.78 (1H, m), 2.01-2.19 (1H,m), 2.47-2.63 (1H, m), 3.36 (3H, s), 3.80-4.06 (2H, m), 4.08-4.24 (1H,m), 4.62 (1H, d, J=6.7 Hz), 4.68 (1H, d, J=6.7 Hz), 4.97-5.19 (4H, m),7.29-7.42 (5H, m).

MS (m/z): 295 (M-Boc+H)⁺.

Step 11 benzyl [(1R,3S,4R)-3-amino-4-hydroxycyclopentyl]carbamatehydrochloride

A mixture of the compound (120 g) obtained in the above Step 10,hydrogen chloride (4 mol/L, 1,4-dioxane solution, 420 mL) and methanol(420 mL) was stirred at room temperature for 30 min. The reactionsolution was concentrated under reduced pressure to give the titlecompound (87.2 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.51-1.70 (2H, m), 2.15-2.29 (2H, m), 3.20-3.35 (1H,m), 3.72-4.14 (3H, m), 5.01 (2H, s), 7.27-7.49 (5H, m), 8.00 (3H, s).

MS (m/z): 251 (M+H)⁻.

Step 12 benzyl{(1R,3R,4S)-3-hydroxy-4-[(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

A mixture of the compound (87.2 g) obtained in the above Step 11,2-nitrobenzenesulfonyl chloride (74.5 g), DIPEA (159 mL) anddichloromethane (1000 mL) was stirred at 0° C. for 1 hr. Water was addedto the reaction solution, and the mixture was extracted withdichloromethane. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure. Ethyl acetate was added to the residue, and the solid wascollected by filtration to give the title compound (33.0 g) as a solid.The filtrate was concentrated, ethyl acetate was added to the residue,and the solid was collected by filtration to give the title compound(25.2 g) as a solid. The filtrate was concentrated, and the residue waspurified by amino silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (40.9 g) as a solid. In total, thetitle compound (99.1 g) was obtained as a solid.

¹H-NMR (CDCl₃) δ: 1.38-1.88 (2H, m), 2.08-2.34 (2H, m), 3.42 (1H, br s),3.59-3.73 (1H, m), 3.86-4.09 (2H, m), 5.05 (2H, s), 5.29 (1H, d, J=6.1Hz), 6.00 (1H, br s), 7.29-7.51 (5H, m), 7.72-7.76 (2H, m), 7.86-7.89(1H, m), 8.13-8.16 (1H, m).

Step 13 benzyl{(1R,3R,4S)-3-hydroxy-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

To a mixture of the compound (87.2 g) obtained in the above Step 12 andDMF (630 mL) were added cesium carbonate (74.2 g) and methyl iodide(21.3 mL) at 0° C., and the mixture was stirred at room temperature for2 hr. Water was added to the reaction solution, and the mixture wasextracted twice with ethyl acetate. The combined organic layers werewashed three times with water and saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure. Ethylacetate/n-hexane was added to the residue, and the solid was collectedby filtration to give the title compound (80.9 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.60-1.69 (1H, m), 1.96-2.09 (1H, m), 2.16-2.33 (2H,m), 2.98-3.11 (4H, m), 3.89-4.06 (2H, m), 4.22-4.30 (1H, m), 5.08 (2H,s), 5.26 (1H, d, J=7.3 Hz), 7.29-7.41 (5H, m), 7.61-7.76 (3H, m),8.02-8.05 (1H, m).

MS (m/z): 450 (M+H)⁻.

Step 14 benzyl[(1R,3R,4S)-3-hydroxy-4-(methylamino)cyclopentyl]carbamate

A mixture of the compound (40.4 g) obtained in the above Step 13, cesiumcarbonate (52.7 g), 4-isopropylbenzenethiol (16.8 mL), THF (140 mL) andmethanol (140 mL) was stirred at room temperature for 3 hr. Amino silicagel was added to the reaction solution, and the mixture was concentratedunder reduced pressure. The residue was purified by amino silica gelcolumn chromatography (n-hexane/ethyl acetate, followed by ethylacetate/methanol) to give an oil. Ethyl acetate/n-hexane was addedthereto, and the solid was collected by filtration to give the titlecompound (22.0 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.28-1.39 (1H, m), 1.78-1.89 (1H, m), 1.91-2.04 (1H,m), 2.33-2.48 (4H, m), 2.78-2.88 (1H, m), 3.95-4.03 (1H, m), 4.11-4.24(1H, m), 5.07 (2H, s), 5.54 (1H, d, J=9.2 Hz), 7.28-7.40 (5H, m).

MS (m/z): 265 (M+H)⁺.

Reference Example A-2 tert-butyl [(1S,3R)-3-aminocyclohexyl]carbamateStep 1 benzyl tert-butyl (1R,3S)-cyclohexane-1,3-diylbiscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using(1R,3S)-3-(tert-butoxycarbonylamino)cyclohexanecarboxylic acid (CAS:222530-34-9).

¹H-NMR (CDCl₃) δ: 0.90-1.07 (3H, m), 1.31-1.42 (1H, m), 1.44 (9H, s),1.72-1.81 (1H, m), 1.93-2.03 (2H, m), 2.26-2.31 (1H, m), 3.42-3.60 (2H,m), 4.39 (1H, br s), 4.58-4.64 (1H, m), 5.08 (2H, s), 7.28-7.72 (5H, m).

MS (m/z): 249 (M−Boc+H)⁺.

Step 2 tert-butyl [(1S,3R)-3-aminocyclohexyl]carbamate

The compound (12.37 g) obtained in the above Step 1 was suspended inethanol (120 mL), 10% palladium on carbon wet (4.0 g) was added thereto,and the mixture was stirred under hydrogen atmosphere at roomtemperature for 4 hr. The palladium catalyst was removed by filtration,and the filtrate was concentrated under reduced pressure to give thetitle compound (8.26 g) as a solid. This was directly used in the nextstep.

¹H-NMR (CDCl₃) δ: 0.88-1.03 (3H, m), 1.28-1.39 (1H, m), 1.44 (9H, s),1.54 (2H, br s), 1.73-1.82 (2H, m), 1.88-1.95 (1H, m), 2.07-2.13 (1H,m), 2.73-2.81 (1H, m), 3.48 (1H, br s), 4.57 (1H, br s).

Reference Example A-3N-[(1R,3S)-3-aminocyclopentyl]-N-methyl-2-nitrobenzene-1-sulfonamideStep 1 benzyl tert-butyl (1R,3S)-cyclopentane-1,3-diylbiscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using(−)-(1R,3S)—N—BOC-3-aminocyclopentanecarboxylic acid (CAS: 161660-94-2).

¹H-NMR (DMSO-D₆) δ: 1.20-1.30 (1H, m), 1.37 (9H, s), 1.43-1.50 (2H, m),1.72-1.79 (2H, m), 2.10-2.17 (1H, m), 3.69-3.81 (2H, m), 5.00 (2H, s),6.85 (1H, d, J=7.3 Hz), 7.29-7.39 (5H, m).

Step 2 tert-butyl{(1S,3R)-3-[(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The compound (19.0 g) obtained in the above Step 1 was suspended inethanol (280 mL), 10% palladium on carbon wet (2.5 g) was added thereto,and the mixture was stirred under hydrogen atmosphere at roomtemperature for 1.5 hr. The palladium was removed by filtration, and thefiltrate was concentrated under reduced pressure. Dichloromethane (230mL) and DIPEA (14.8 mL) were added thereto, and 2-nitrobenzenesulfonylchloride (12.6 g) was added thereto under ice-cooling. After stirring atroom temperature for 2 hr, dichloromethane and water were added to thereaction solution, and the mixture was subjected to liquid separation.The organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (12.77 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.40-1.48 (1H, m), 1.42 (9H, s), 1.55-1.70 (2H, m),1.81-1.99 (2H, m), 2.20-2.27 (1H, m), 3.74-3.82 (1H, m), 3.83-3.93 (1H,m), 4.60 (1H, br s), 5.61 (1H, br s), 7.73-7.78 (2H, m), 7.85-7.87 (1H,m), 8.15-8.17 (1H, m).

[0535]

Step 3 tert-butyl{(1S,3R)-3-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.38-1.51 (2H, m), 1.42 (9H, s), 1.56-2.02 (3H, m),2.14-2.22 (1H, m), 2.85 (3H, s), 3.81-3.89 (1H, m), 4.26-4.34 (1H, m),4.50 (1H, br s), 7.60-7.64 (1H, m), 7.66-7.72 (2H, m), 8.00-8.04 (1H,m).

Step 4N—[(R,3S)-3-aminocyclopentyl]-N-methyl-2-nitrobenzene-1-sulfonamide

The compound (2.04 g) obtained in the above Step 3 was dissolved indichloromethane (20 mL), hydrogen chloride (4 mol/L, 1,4-dioxanesolution, 20 mL) was added thereto, and the mixture was stirred at roomtemperature for 1 hr. The solvent was evaporated under reduced pressure,dichloromethane and saturated aqueous sodium hydrogencarbonate solutionwere added to the residue, and the mixture was subjected to liquidseparation. The aqueous layer was extracted with dichloromethane. Theorganic layers were combined, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby amino silica gel column chromatography (ethyl acetate/methanol) togive the title compound (1.21 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.17-1.40 (4H, m), 1.73-1.86 (3H, m), 2.02-2.09 (1H,m), 2.88 (3H, s), 3.29-3.36 (1H, m), 4.30-4.39 (1H, m), 7.60-7.62 (1H,m), 7.66-7.71 (2H, m), 8.00-8.04 (1H, m).

Reference Example A-4 tert-butyl[(1S,3R)-3-(methylamino)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 14 ofReference Example A-1, using the compound obtained in Step 3 ofReference Example A-3.

¹H-NMR (CDCl₃) δ: 1.33-1.72 (4H, m), 1.44 (9H, s), 1.77-2.11 (3H, m),2.38 (3H, s), 3.03-3.09 (1H, m), 4.00-4.08 (1H, m), 5.30 (1H, br s).

Reference Example A-5 tert-butyl[(1S,3R)-3-(ethylamino)cyclopentyl]carbamate Step 1 tert-butyl{(1S,3R)-3-[ethyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1 except that bromoethane (CAS: 74-96-4) was usedinstead of iodomethane, using the compound obtained in Step 2 ofReference Example A-3.

¹H-NMR (CDCl₃) δ: 1.24 (3H, t, J=7.4 Hz), 1.41-1.52 (2H, m), 1.43 (9H,s), 1.65-1.74 (1H, m), 1.83-2.01 (2H, m), 2.22-2.28 (1H, m), 3.31-3.37(2H, m), 3.81-3.88 (1H, m), 4.07-4.17 (1H, m), 4.54 (1H, br s),7.59-7.63 (1H, m), 7.65-7.72 (2H, m), 7.99-8.03 (1H, m).

MS (m/z): 314 (M−Boc+H)⁺.

Step 2 tert-butyl [(1S,3R)-3-(ethylamino)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 14 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 0.88 (1H, br s), 1.09 (3H, t, J=7.3 Hz), 1.30-1.36(1H, m), 1.44 (9H, s), 1.45-1.65 (2H, m), 1.80-2.09 (3H, m), 2.60 (2H,q, J=7.3 Hz), 3.13-3.19 (1H, m), 4.01 (1H, br s), 5.27 (1H, br s).

MS (m/z): 229 (M+H)⁻.

Reference Example A-6 benzyl(1R,3S,5R)-3-amino-5-(methoxymethoxy)cyclohexane-1-carboxylate(racemate) Step 1 methoxymethyl(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-hydroxycyclohexane-1-carboxylate(racemate)

A mixture of(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-hydroxycyclohexane-1-carboxylicacid (racemate) (11.4 g) synthesized according to the method describedin a literature (Bioorg. Med. Chem. 2006, 14, 2242-2252), DIPEA (20.4mL) and dichloromethane (390 mL) was ice-cooled, chloromethyl methylether (3.08 mL) was added thereto, and the mixture was stirred at roomtemperature for 3 hr. Aqueous saturated ammonium chloride solution wasadded to the reaction solution, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated brine, dried overanhydrous sodium sulfate, and concentrated under reduced pressure togive the title compound (11.8 g) as an oil. The obtained compound wasdirectly used in the next step without purification.

MS (m/z): 338 (M+H)⁺.

Step 2 methoxymethyl(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-(methoxymethoxy)cyclohexane-1-carboxylate(racemate)

A mixture of the compound (11.8 g) obtained in the above Step 1, DIPEA(18.2 mL), chloromethyl methyl ether (3.4 mL) and dichloromethane (350mL) was stirred at room temperature for 2 hr. The reaction solution wasice-cooled, chloromethyl methyl ether (10.5 mL) was added thereto, andthe mixture was stirred at room temperature for additional 18 hr.Aqueous saturated ammonium chloride solution was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate) togive the title compound (7.88 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.24-1.48 (3H, m), 2.29-2.32 (3H, m), 2.44-2.47 (1H,m), 3.36 (3H, s), 3.46 (3H, s), 3.60-3.63 (2H, m), 4.66-4.68 (3H, m),5.11 (2H, s), 5.24 (2H, s), 7.32-7.35 (5H, m).

MS (m/z): 382 (M+H)⁺.

Step 3(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-(methoxymethoxy)cyclohexane-1-carboxylicacid (racemate)

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound (7.88 g) obtained in the aboveStep 2.

¹H-NMR (DMSO-D₆) δ: 1.09-1.24 (3H, m), 1.94-2.11 (3H, m), 2.31-2.34 (1H,m), 3.25 (3H, s), 3.40-3.43 (1H, m), 3.50-3.58 (1H, m), 4.60 (2H, s),5.02 (2H, s), 7.01 (1H, s), 7.27-7.37 (5H, m), 11.88 (1H, br s).

MS (m/z): 338 (M+H)⁺.

Step 4 benzyl prop-2-en-1-yl[(1R,3S,5S)-5-(methoxymethoxy)cyclohexane-1,3-diyl]biscarbamate(racemate)

A mixture of the compound (6.65 g) obtained in the above Step 3,diphenylphosphoryl azide (5.52 mL), TEA (3.57 mL) and toluene (98.6 mL)was stirred at 90° C. for 30 min. Allyl alcohol (1.40 g) was addedthereto, and the mixture was stirred at the same temperature for 5 hr,and concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (6.2 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.03-1.27 (3H, m), 2.18-2.28 (3H, m), 3.34 (3H, s),3.62-3.66 (3H, m), 4.54 (2H, d, J=5.2 Hz), 4.67-4.75 (4H, m), 5.09 (2H,s), 5.20-5.28 (2H, m), 5.87-5.93 (1H, m), 7.30-7.33 (5H, m).

MS (m/z): 393 (M+H)⁺.

Step 5 benzyl(1R,3S,5R)-3-amino-5-(methoxymethoxy)cyclohexane-1-carboxylate(racemate)

A mixture of the compound (335 mg) obtained in the above Step 4,dichloromethane (8 mL), pyrrolidine (0.177 mL) andtetrakis(triphenylphosphine)palladium(0) (20 mg) was stirred at roomtemperature for 30 min, and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography (ethylacetate/methanol) to give the title compound (93 mg) as a solid.

¹H-NMR (CDCl₃) δ: 0.93-0.95 (1H, m), 1.07-1.13 (2H, m), 2.12-2.18 (2H,m), 2.29-2.32 (1H, m), 2.78-2.81 (1H, m), 3.35 (3H, s), 3.57-3.62 (2H,m), 4.61-4.65 (3H, m), 5.10 (2H, s), 7.28-7.32 (5H, m).

MS (m/z): 309 (M+H)⁺.

Reference Example A-7 benzyl[(1S,3R,5S)-3-amino-5-(dimethylcarbamoyl)cyclohexyl]carbamate (racemate)Step 1 benzyl[(1S,3R,5R)-3-(dimethylcarbamoyl)-5-(methoxymethoxy)cyclohexyl]carbamate(racemate)

To a mixture of the compound (1.59 g) obtained in Step 3 of ReferenceExample A-6 and dichloromethane (15 mL) were added dimethylaminehydrochloride (CAS: 506-59-2) (0.48 g), anhydrous HOBt (CAS:123333-53-9) (0.77 g), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (CAS: 1892-57-5) (1.10 g) and TEA (1.31 mL) at roomtemperature. This reaction solution was warmed to room temperature, andstirred for 18 hr. Ethyl acetate (30 mL) was added to this reactionsolution, and the mixture was washed successively with saturated aqueousammonium chloride solution (10 mL), saturated aqueous sodiumhydrogencarbonate solution (10 mL) and saturated brine (10 mL), and theorganic layer was dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (methanol/dichloromethane) to give the title compound(1.65 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.21 (1H, q, J=11.7 Hz), 1.36 (1H, q, J=12.5 Hz), 1.51(1H, q J=12.5 Hz), 2.02 (2H, t, J=14.0 Hz), 2.35-2.36 (1H, m), 2.64-2.70(1H, m), 2.95 (3H, s), 3.08 (3H, s), 3.38 (3H, s), 3.64-3.67 (2H, m),4.67-4.71 (2H, m), 4.74-4.76 (1H, m), 5.11 (2H, s), 7.33-7.41 (5H, m).

MS (m/z): 365 (M+H)⁺.

Step 2 benzyl[(1S,3R,5R)-3-(dimethylcarbamoyl)-5-hydroxycyclohexyl]carbamate(racemate)

To a mixture of the compound (1.65 g) obtained in the above Step 1 and1,4-dioxane (5 mL) was added hydrogen chloride (4 mol/L, 1,4-dioxanesolution, 5.66 mL) at room temperature. This reaction solution wasstirred at room temperature for 2 hr, and water (20 mL) and ethylacetate (30 mL) were added thereto. This mixture was washed twice withwater (10 mL), and washed successively with saturated aqueous sodiumhydrogencarbonate solution (10 mL) and saturated brine (20 mL), and theorganic layer was dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (methanol/dichloromethane) to give the title compound(1.12 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.25 (1H, q, J=11.5 Hz), 1.46 (1H, q, J=12.1 Hz),1.52-1.61 (1H, m), 1.99-2.02 (2H, m), 2.29-2.30 (2H, m), 2.69-2.75 (1H,m), 2.95 (3H, s), 3.07 (3H, s), 3.69-3.72 (1H, m), 3.74-3.82 (1H, m),5.01 (1H, d, J=7.9 Hz), 5.10 (2H, s), 7.34-7.39 (5H, m).

Step 3(1S,3S,5R)-3-{[(benzyloxy)carbonyl]amino}-5-(dimethylcarbamoyl)cyclohexyl4-nitrobenzoate (racemate)

To a mixture of the compound (1.06 g) obtained in the above Step 2,4-nitrobenzoic acid (CAS: 62-23-7) (0.66 g), triphenylphosphine (CAS:14264-16-5) (purity 97%, 1.04 g) and THF (15 mL) was added diisopropylazodicarboxylate (40% toluene solution) (CAS: 2446-83-5) (2.1 mL) underice-cooling. This mixture was warmed to room temperature, and stirred atroom temperature for 5.5 hr. Saturated aqueous sodium hydrogencarbonatesolution (20 mL), ethyl acetate (20 mL) and saturated brine (20 mL) wereadded thereto, and the mixture was subjected to liquid separation. Theorganic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (methanol/dichloromethane)to give the title compound (1.55 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.53-1.58 (2H, m), 1.90-1.95 (1H, m), 2.10-2.11 (1H,m), 2.20-2.23 (1H, m), 2.35-2.39 (1H, m), 2.97 (3H, s), 3.06 (3H, s),3.09-3.17 (1H, m), 4.06-4.10 (1H, m), 4.87-4.89 (1H, m), 5.12 (2H, s),5.60 (1H, s), 7.37-7.38 (5H, m), 8.23 (2H, d, J=8.8 Hz), 8.34 (2H, d,J=8.8 Hz).

MS (m/z): 470 (M+H)⁺.

Step 4 benzyl[(1S,3R,5S)-3-(dimethylcarbamoyl)-5-hydroxycyclohexyl]carbamate(racemate)

To a mixture of the compound (1.55 g) obtained in the above Step 3 andethanol (22 mL) was added potassium carbonate (CAS: 584-08-7) (1.37 g)at room temperature, and the mixture was stirred for 2 hr. Saturatedaqueous sodium hydrogencarbonate solution (20 mL), ethyl acetate (20 mL)and saturated brine (20 mL) were added thereto, and the mixture wassubjected to liquid separation. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (methanol/dichloromethane) to give the title compound(0.710 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.36-1.44 (1H, m), 1.44-1.50 (1H, m), 1.64-1.71 (1H,m), 1.81-1.84 (1H, m), 2.06-2.10 (3H, m), 2.95 (3H, s), 3.09 (3H, s),3.22-3.26 (1H, m), 4.03-4.11 (1H, m), 4.32 (1H, s), 4.92 (1H, d, J=8.5Hz), 5.10 (2H, s), 7.34-7.37 (5H, m).

MS (m/z): 321 (M+H)⁺.

Step 5 benzyl[(1S,3R,5R)-3-azido-5-(dimethylcarbamoyl)cyclohexyl]carbamate (racemate)

To a mixture of the compound (0.513 g) obtained in the above Step 4,triphenylphosphine (CAS: 14264-16-5) (0.752 g), diphenylphosphoryl azide(0.412 mL) and THF (15 mL) was added diisopropyl azodicarboxylate (40%toluene solution) (1.50 mL) under ice-cooling. This reaction solutionwas warmed to room temperature, and stirred for 3 hr. Ethyl acetate (30mL) was added to this reaction solution, and the mixture was washedtwice with saturated brine (10 mL). The organic layer was dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography(methanol/dichloromethane) to give the title compound (0.553 g) as anoil.

¹H-NMR (CDCl₃) δ: 1.17-1.27 (1H, m), 1.39 (1H, q, J=12.2 Hz), 1.57 (1H,q, J=12.5 Hz), 1.99-2.02 (2H, m), 2.33-2.35 (1H, m), 2.68-2.74 (1H, m),2.94 (3H, s), 3.06 (3H, s), 3.38-3.44 (1H, m), 3.67-3.71 (1H, m), 5.10(2H, s), 6.55 (1H, s), 7.33-7.38 (5H, m).

MS (m/z): 346 (M+H)⁺.

Step 6 benzyl[(1S,3R,5S)-3-amino-5-(dimethylcarbamoyl)cyclohexyl]carbamate (racemate)

To a mixture of the compound (0.553 g) obtained in the above Step 5 andTHF (15 mL) was added triphenylphosphine (CAS: 14264-16-5) (0.840 g)under ice-cooling. This reaction solution was warmed to roomtemperature, and stirred for 30 min. Water (10 mL) was added to thisreaction solution, and the mixture was allowed to stand for 19 hr,heated to 85° C., and stirred for 2 hr. The reaction solution wasconcentrated under reduced pressure. Ethanol (20 mL) was added to theresidue, and the mixture was concentrated under reduced pressure. Theseconcentration operations were conducted three times to give the crudetitle compound (0.511 g) as an oil. The product was used in the nextstep without purification.

MS (m/z): 320 (M+H)⁺.

Reference Example A-8 benzyl [(1S,3R)-3-aminocyclohexyl]carbamatehydrochloride Step 1 benzyl tert-butyl(1R,3S)-cyclohexane-1,3-diylbiscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using(1S,3R)-3-[(tert-butoxycarbonyl)amino]cyclohexane-1-carboxylic acid(CAS: 222530-34-9).

¹H-NMR (CDCl₃)b: 1.05-0.87 (3H, m), 1.47-1.33 (10H, m), 1.82-1.73 (1H,m), 2.04-1.91 (2H, m), 2.33-2.25 (1H, m), 3.66-3.38 (2H, m), 4.38 (1H,br s), 4.59 (1H, br s), 5.08 (2H, br s), 7.40-7.28 (5H, m).

Step 2 benzyl [(1S,3R)-3-aminocyclohexyl]carbamate hydrochloride

The compound (1.90 g) obtained in the above Step 1 was dissolved inhydrogen chloride (4 mol/L, 1,4-dioxane solution, 54.5 mL), and thesolution was stirred at room temperature for 30 min. The reactionsolution was concentrated under reduced pressure, and the residue wassubjected to slurry washing with ethyl acetate to give the titlecompound (1.54 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 0.97-1.37 (4H, m), 1.68-1.90 (3H, m), 2.09 (1H, d,J=11.5 Hz), 2.96-3.12 (1H, m), 3.32-3.42 (1H, m), 5.01 (2H, s),7.29-7.42 (5H, m), 7.99 (3H, br s).

Reference Example A-9 methyl(1S,3R,5S)-3-amino-5-{[(benzyloxy)carbonyl]amino}cyclohexane-1-carboxylate(racemate) Step 1(1S,3S,5R)-3-{[(benzyloxy)carbonyl]amino}-5-(methoxycarbonyl)cyclohexyl4-nitrobenzoate (racemate)

The title compound was obtained in the same manner as in Step 3 ofReference Example A-7, using methyl(1R,3S,5R)-3-(benzyloxycarbonylamino)-5-hydroxy-cyclohexanecarboxylate(racemate) synthesized according to the method described in a literature(Bioorg. Med. Chem., 2006, 14, 2242-2252).

MS (m/z): 457 (M+H)⁺.

Step 2 methyl(1R,3S,5S)-3-{[(benzyloxy)carbonyl]amino}-5-hydroxycyclohexane-1-carboxylate(racemate)

The title compound was obtained in the same manner as in Step 4 ofReference Example A-7, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.24-1.39 (2H, m), 1.51-1.62 (2H, m), 1.97-2.12 (2H,m), 2.25-2.34 (1H, m), 2.86-3.02 (1H, m), 3.67 (3H, s), 3.94-4.08 (1H,m), 4.26-4.33 (1H, m), 4.60-4.74 (1H, m), 5.08 (2H, s), 7.28-7.39 (5H,m).

MS (m/z): 308 (M+H)⁺.

Step 3 methyl(1R,3R,5S)-3-azido-5-{[(benzyloxy)carbonyl]amino}cyclohexane-1-carboxylate(racemate)

The title compound was obtained in the same manner as in Step 5 ofReference Example A-7, using the compound obtained in the above Step 2.

MS (m/z): 333 (M+H)⁺.

Step 4 methyl(1S,3R,5S)-3-amino-5-{[(benzyloxy)carbonyl]amino}cyclohexane-1-carboxylate(racemate)

The title compound was obtained in the same manner as in Step 6 ofReference Example A-7, using the compound obtained in the above Step 3.

¹H-NMR (CDCl₃) δ: 0.90-1.79 (5H, m), 2.06-2.20 (2H, m), 2.22-2.30 (1H,m), 2.41-2.53 (1H, m), 2.77-2.90 (1H, m), 3.53-3.70 (1H, m), 3.68 (3H,s), 4.63-4.75 (1H, m), 5.09 (2H, s), 7.29-7.41 (5H, m).

MS (m/z): 307 (M+H)⁺.

Reference Example A-10 tert-butyl[(1S,2S,5S)-5-amino-2-methoxycyclohexyl]carbamate Step 1 ethyl(1R,3S,4S)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclohexane-1-carboxylate

A mixture ofethyl(R,3S,4S)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclohexane-1-carboxylate(13.84 g) synthesized according to the method described in a literature(Tetrahedron 2017, 73, 1381-1388), dimethoxyethane (155 mL), sodiumiodide (CAS: 7681-82-5) (6.97 g), chloromethyl methyl ether (10.5 mL)and DIPEA (49 mL) was heated under reflux at 105° C. for 2 hr. Saturatedbrine and water were added to the reaction solution, the mixture wasextracted with ethyl acetate, and the organic layer was dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography(dichloromethane/ethyl acetate) to give the title compound (14.43 g) asa solid.

¹H-NMR (CDCl₃) δ: 1.26 (3H, t, J=7.0 Hz), 1.45 (9H, s), 1.57-1.78 (4H,m), 1.89-2.00 (1H, m), 2.21-2.30 (1H, m), 2.43-2.52 (1H, m), 3.39 (3H,s), 3.51-3.59 (1H, m), 3.68-3.80 (1H, m), 4.15 (2H, q, J=7.0 Hz), 4.60(1H, br s), 4.66 (1H, d, J=6.7 Hz), 4.69 (1H, d, J=6.7 Hz).

Step 2(1S,3S,4S)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclohexane-1-carboxylicacid

The compound (14.43 g) synthesized in the above Step 1 was dissolved inethanol (150 mL), sodium ethoxide (CAS: 141-52-6) (concentration 20%,ethanol solution, 24 mL) was added thereto, and the mixture was stirredat 50° C. for 40 hr. 1N Hydrochloric acid ethanol solution (CAS:7647-01-0) (61 mL) was added to the reaction solution, and the mixturewas concentrated under reduced pressure. Water was added to the obtainedresidue, and the mixture was extracted with dichloromethane. The organiclayer was dried over anhydrous sodium sulfate, and concentrated underreduced pressure. The residue was subjected to silica gel columnchromatography (dichloromethane/ethyl acetate). The obtained residue wassuspended in n-hexane/ethyl acetate, and the solid was collected byfiltration to give a mixture (4.11 g) containing the title compound as asolid.

Step 3 benzyl tert-butyl[(1S,3S,4S)-4-(methoxymethoxy)cyclohexane-1,3-diyl]biscarbamate

A mixture of the compound (4.11 g) obtained in the above Step 2,1,4-dioxane (66 mL), diphenylphosphoryl azide (CAS: 26386-88-9) (3.5 mL)and TEA (2.5 mL) was stirred at 90° C. for 2 hr. Then, benzyl alcohol(CAS: 100-51-6) (2.2 mL) was added thereto, and the mixture was stirredat 90° C. for 5 hr. Saturated brine and water were added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was dried over anhydrous sodium sulfate, and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (dichloromethane/ethyl acetate) to give the titlecompound (2.64 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.08-1.25 (2H, m), 1.40-1.46 (1H, m), 1.43 (9H, s),2.00-2.13 (2H, m), 2.37-2.46 (1H, m), 3.21-3.32 (1H, m), 3.39-3.51 (1H,m), 3.39 (3H, s), 3.53-3.66 (1H, m), 4.59 (1H, br s), 4.60 (1H, d, J=6.7Hz), 4.70 (1H, d, J=6.7 Hz), 4.74 (1H, br s), 5.08 (2H, s), 7.29-7.40(5H, m).

Step 4 benzyl tert-butyl[(1S,3S,4S)-4-hydroxycyclohexane-1,3-diyl]biscarbamate

The compound (0.595 g) obtained in the above Step 3 was dissolved in1,4-dioxane (18 mL), hydrogen chloride (4 mol/L, 1,4-dioxane solution,36 mL) was added thereto, and the mixture was stirred at roomtemperature for 3 hr. Then, the solvent was evaporated under reducedpressure, and the residue was dried. A solution of THF (10 mL), TEA(0.57 mL) and di-tert-butyl dicarbonate (CAS: 24424-99-5) (0.418 g) inTHF (6.0 mL) was added to the obtained residue, and the mixture wasstirred at room temperature for 2.5 hr. Saturated aqueous sodiumbicarbonate solution was added to the reaction solution, and the mixturewas extracted with dichloromethane. The organic layer was dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (0.403 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.06-1.24 (2H, m), 1.37-1.49 (1H, m), 1.45 (9H, s),1.96-2.10 (2H, m), 2.24-2.32 (1H, m), 3.26-3.50 (3H, m), 3.53-3.68 (1H,m), 4.53 (1H, br s), 4.59 (1H, d, J=6.1 Hz), 5.08 (2H, s), 7.30-7.39(5H, m).

Step 5 benzyl tert-butyl[(1S,3S,4S)-4-methoxycyclohexane-1,3-diyl]biscarbamate

A mixture of the compound (0.350 g) obtained in the above Step 4,dichloromethane (14 mL), silver(I) oxide (CAS: 20667-12-3) (0.677 g),molecular sieves 3A (CAS: 308080-99-1) (0.460 g, used after dried underreduced pressure at 180° C. for 2 hr) and methyl iodide (CAS: 74-88-4)(1.2 mL) was stirred vigorously at 45° C. for 9 hr. The reactionsolution was filtered through Celite, and the filtrate was concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (n-hexane/ethyl acetate) to give the titlecompound (0.269 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.08-1.38 (3H, m), 1.44 (9H, s), 2.03-2.20 (2H, m),2.41-2.50 (1H, m), 2.94-3.07 (1H, m), 3.32-3.44 (1H, m), 3.36 (3H, s),3.52-3.66 (1H, m), 4.50-4.67 (2H, m), 5.08 (2H, s), 7.29-7.39 (5H, m).

MS (m/z): 279 (M−Boc+H)⁺.

Step 6 tert-butyl [(1S,2S,5S)-5-amino-2-methoxycyclohexyl]carbamate

A mixture of the compound (0.267 g) obtained in the above Step 5, 5%palladium on carbon (PH) wet (CAS: 7440-05-3) (0.304 g) and ethanol (12mL) was stirred under hydrogen atmosphere at room temperature for 3.5hr. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure. The obtained residue was purifiedby amino silica gel column chromatography (ethyl acetate/methanol) togive the title compound (0.161 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.11-1.22 (2H, m), 1.27-1.40 (1H, m), 1.45 (9H, s),1.83-1.92 (1H, m), 2.07-2.15 (1H, m), 2.17-2.27 (1H, m), 2.92-3.00 (1H,m), 3.04 (1H, td, J=8.7, 3.9 Hz), 3.37 (3H, s), 3.46-3.56 (1H, m), 5.25(1H, br s).

Reference Example A-11 benzyl[(1R,3S,5S)-3-amino-5-methoxycyclohexyl]carbamate (racemate) Step 1methyl(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-methoxycyclohexane-1-carboxylate(racemate)

To a solution of(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-hydroxycyclohexane-1-carboxylicacid (racemate) (0.498 g) synthesized according to the method describedin a literature (Bioorg. Med. Chem. 2006, 14, 2242-2252) anddichloromethane (17.0 mL) were added silver(I) oxide (1.57 g), molecularsieves 3A (0.249 g) and methyl iodide (2.11 mL), and the mixture wasstirred at 40° C. for 3 hr. Additional methyl iodide (2.11 mL) was addedthereto, and the mixture was stirred at 40° C. for 7.5 hr. The insolublesubstance was removed by filtration through Celite, and washed withdichloromethane and methanol, and the filtrate was concentrated underreduced pressure. The obtained crude product was purified by columnchromatography (methanol/dichloromethane) to give the title compound(0.480 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.06 (1H, q, J=11.5 Hz), 1.19-1.32 (2H, br m),2.20-2.41 (4H, br m), 3.17-3.26 (1H, m), 3.34 (3H, s), 3.55-3.62 (1H,m), 3.66 (3H, s), 5.05-5.14 (1H, m), 5.07 (2H, s), 7.32-7.34 (5H, br m).

MS (m/z): 322 (M+H)⁺.

Step 2(1S,3R,5S)-3-{[(benzyloxy)carbonyl]amino}-5-methoxycyclohexane-1-carboxylicacid (racemate)

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound obtained in the above Step 1.The obtained crude product was directly used in the next step.

Step 3 benzyl prop-2-en-1-yl[(1R,3S,5S)-5-methoxycyclohexane-1,3-diyl]biscarbamate (racemate)

The title compound was obtained in the same manner as in Step 4 ofReference Example A-6, using the compound obtained in the above Step 2.

MS (m/z): 363 (M+H)⁺.

Step 4 benzyl [(1R,3S,5S)-3-amino-5-methoxycyclohexyl]carbamate(racemate)

The title compound was obtained in the same manner as in Step 5 ofReference Example A-6, using the compound obtained in the above Step 3.

¹H-NMR (CDCl₃) δ: 0.90-1.03 (3H, m), 1.35 (2H, s), 2.12 (1H, d, J=11.6Hz), 2.21 (1H, d, J=12.2 Hz), 2.36 (1H, d, J=11.6 Hz), 2.79 (1H, t,J=11.3 Hz), 3.24 (1H, t, J=10.7 Hz), 3.35 (3H, s), 3.58 (1H, d, J=7.9Hz), 4.72 (1H, s), 5.09 (2H, s), 7.30-7.36 (5H, m).

MS (m/z): 279 (M+H)⁺.

Reference Example A-12 tert-butyl[(1R,2R,4S)-2-(methoxymethoxy)-4-(methylamino)cyclopentyl]carbamate Step1 methyl(1S,3R,4R)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylate

The title compound was obtained in the same manner as in Step 2 ofReference Example A-6, using methyl(1S,3R,4R)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclopentane-1-carboxylate(CAS: 321744-16-5).

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.67-1.75 (1H, m), 2.05-2.13 (2H, m),2.40 (1H, br s), 3.00-3.08 (1H, m), 3.37 (3H, s), 3.70 (3H, s), 3.97(1H, br s), 4.00-4.04 (1H, m), 4.63 (1H, d, J=6.7 Hz), 4.72 (1H, d,J=6.7 Hz), 5.10 (1H, br s).

MS (m/z): 204 (M−Boc+H)⁺.

Step 2(1S,3R,4R)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylicacid

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound obtained in the above Step 1.

Step 3 benzyl tert-butyl[(1S,3R,4R)-4-(methoxymethoxy)cyclopentane-1,3-diyl]biscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.37-1.62 (1H, m), 1.44 (9H, s), 1.93-2.09 (2H, m),2.51-2.61 (1H, m), 3.35 (3H, s), 3.73-3.81 (1H, m), 4.01-4.17 (2H, m),4.62 (1H, d, J=6.7 Hz), 4.69 (1H, d, J=6.7 Hz), 5.08-5.23 (4H, m),7.30-7.40 (5H, m).

Step 4 tert-butyl[(1R,2R,4S)-4-amino-2-(methoxymethoxy)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 2 ofReference Example A-2, using the compound obtained in the above Step 3.

¹H-NMR (DMSO-D₆) δ: 1.10-1.20 (1H, m), 1.37 (9H, s), 1.53-1.61 (1H, m),1.69-1.77 (1H, m), 2.03-2.11 (1H, m), 3.21 (3H, s), 3.24-3.32 (1H, m),3.58-3.70 (1H, m), 3.81-3.91 (1H, m), 4.49-4.55 (1H, m), 4.57-4.62 (1H,m), 6.91-6.94 (1H, m).

Step 5 tert-butyl{(1R,2R,4S)-2-(methoxymethoxy)-4-[(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 12 ofReference Example A-1, using the compound obtained in the above Step 4.

¹H-NMR (DMSO-D₆) δ: 1.24-1.34 (1H, m), 1.36 (9H, s), 1.66-1.79 (2H, m),2.05-2.17 (1H, m), 3.17 (3H, s), 3.52-3.63 (1H, m), 3.65-3.75 (1H, m),3.75-3.82 (1H, m), 4.47 (1H, d, J=6.7 Hz), 4.54 (1H, d, J=6.7 Hz), 6.96(1H, d, J=7.9 Hz), 7.84-7.93 (2H, m), 7.95-8.01 (1H, m), 8.02-8.07 (1H,m), 8.09-8.15 (1H, m).

Step 6 tert-butyl{(1R,2R,4S)-2-(methoxymethoxy)-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1, using the compound obtained in the above Step 5.

¹H-NMR (DMSO-D₆) δ: 1.36 (9H, s), 1.42-1.52 (1H, m), 1.59-1.67 (1H, m),1.82-1.96 (2H, m), 2.79 (3H, s), 3.20 (3H, s), 3.60-3.68 (1H, m),3.76-3.81 (1H, m), 4.31-4.40 (1H, m), 4.52 (1H, d, J=6.7 Hz), 4.59 (1H,d, J=6.7 Hz), 7.00 (1H, d, J=7.9 Hz), 7.83-7.92 (2H, m), 7.96-8.03 (2H,m).

Step 7 tert-butyl[(1R,2R,4S)-2-(methoxymethoxy)-4-(methylamino)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 14 ofReference Example A-1, using the compound obtained in the above Step 6.

¹H-NMR (DMSO-D₆) δ: 1.16-1.26 (1H, m), 1.37 (9H, s), 1.59-1.68 (1H, m),1.69-1.78 (1H, m), 2.03-2.12 (1H, m), 2.20 (3H, s), 2.94-3.01 (1H, m),3.22 (3H, s), 3.63-3.70 (1H, m), 3.82-3.86 (1H, m), 4.52 (1H, d, J=6.1Hz), 4.59 (1H, d, J=6.7 Hz), 6.82-6.87 (1H, m).

Reference Example A-13 tert-butyl[(1R,2S,4S)-2-(methoxymethoxy)-4-(methylamino)cyclopentyl]carbamate Step1 methyl(1S,3R,4S)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylate

The title compound was obtained in the same manner as in Step 2 ofReference Example A-6, using methyl(1R,2S,4S)—N—BOC-1-amino-2-hydroxycyclopentane-4-carboxylate (CAS:321744-14-3).

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.84-1.94 (1H, m), 2.05-2.33 (3H, m),2.75-2.85 (1H, m), 3.37 (3H, s), 3.68 (3H, s), 3.90-3.99 (1H, m),4.00-4.05 (1H, m), 4.61 (1H, d, J=6.7 Hz), 4.69 (1H, d, J=6.7 Hz),5.04-5.12 (1H, m).

Step 2(1S,3R,4S)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylicacid

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound obtained in the above Step 1.

Step 3 benzyl tert-butyl[(1S,3R,4S)-4-(methoxymethoxy)cyclopentane-1,3-diyl]biscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.39-1.48 (1H, m), 1.44 (9H, s), 1.73-1.80 (1H, m),2.08-2.16 (1H, m), 2.52-2.60 (1H, m), 3.37 (3H, s), 3.85-3.97 (1H, m),4.00-4.05 (1H, m), 4.12-4.22 (1H, m), 4.61-4.71 (2H, m), 5.01-5.13 (4H,m), 7.30-7.41 (5H, m).

Step 4 tert-butyl[(1R,2S,4S)-4-amino-2-(methoxymethoxy)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 2 ofReference Example A-2, using the compound obtained in the above Step 3.

Step 5 tert-butyl{(1R,2S,4S)-2-(methoxymethoxy)-4-[(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 12 ofReference Example A-1, using the compound obtained in the above Step 4.

¹H-NMR (CDCl₃) δ: 1.35-1.44 (1H, m), 1.41 (9H, s), 1.77-1.82 (1H, m),1.91-2.00 (1H, m), 2.32-2.40 (1H, m), 3.39 (3H, s), 3.82-3.90 (1H, m),3.98-4.03 (2H, m), 4.65 (1H, d, J=6.7 Hz), 4.68 (1H, d, J=6.7 Hz),4.97-5.06 (1H, m), 5.88 (1H, d, J=9.7 Hz), 7.72-7.78 (2H, m), 7.85-7.90(1H, m), 8.11-8.16 (1H, m).

Step 6 tert-butyl{(1R,2S,4S)-2-(methoxymethoxy)-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1, using the compound obtained in the above Step 5.

¹H-NMR (CDCl₃) δ: 1.43 (9H, s), 1.65-1.76 (2H, m), 2.06-2.12 (1H, m),2.17-2.25 (1H, m), 2.86 (3H, s), 3.37 (3H, s), 3.77-3.87 (1H, m),3.95-3.99 (1H, m), 4.35-4.45 (1H, m), 4.62 (1H, d, J=6.7 Hz), 4.69 (1H,d, J=6.7 Hz), 4.98-5.06 (1H, m), 7.60-7.65 (1H, m), 7.67-7.74 (2H, m),8.01-8.04 (1H, m).

Step 7 tert-butyl[(1R,2S,4S)-2-(methoxymethoxy)-4-(methylamino)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 14 ofReference Example A-1, using the compound obtained in the above Step 6.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.51-1.64 (2H, m), 2.04-2.15 (1H, m),2.24-2.33 (1H, m), 2.37 (3H, s), 2.98-3.05 (1H, m), 3.38 (3H, s),3.88-3.99 (1H, m), 4.00-4.06 (1H, m), 4.64 (1H, d, J=6.7 Hz), 4.68 (1H,d, J=6.7 Hz), 5.19-5.26 (1H, m).

Reference Example A-14 benzyl[(1R,3S,4S)-3-hydroxy-4-(methylamino)cyclopentyl]carbamate Step 1 methyl(1R,3S,4S)-3-(acetyloxy)-4-[(tert-butoxycarbonyl)amino]cyclopentane-1-carboxylate

Methyl(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclopentane-1-carboxylate(CAS: 321744-23-4) (0.300 g) was dissolved in THF (2.3 mL), anddiphenyl-2-pyridylphosphine (0.487 g), 1,1′-(azodicarbonyl)dipiperazine(0.394 g) and acetic acid (0.132 mL) were added thereto, and the mixturewas stirred at room temperature for 2 hr. 2N Hydrochloric acid and ethylacetate were added to the reaction solution, and the mixture wassubjected to extraction operation. The organic layer was washed withsaturated aqueous sodium bicarbonate solution and saturated brine, anddried over anhydrous sodium sulfate. The solvent was evaporated underreduced pressure, and the residue was washed with a mixed solvent ofn-hexane/ethyl acetate=3/1, whereby the insoluble substance was removedby filtration. The filtrate was concentrated under reduced pressure, andthe residue was purified by silica gel chromatography (ethylacetate/dichloromethane) to give the title compound (0.230 g) as asolid.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.76 (1H, dt, J=14.0, 6.0 Hz), 1.98-2.08(4H, m), 2.21-2.29 (1H, m), 2.38-2.52 (1H, m), 2.96-3.05 (1H, m), 3.71(3H, s), 4.01 (1H, br s), 5.01-5.15 (2H, m). Step 2(1R,3S,4S)-3-[(tert-butoxycarbonyl)amino]-4-hydroxycyclopentane-1-carboxylicacid

The compound (46.6 g) obtained in the above Step 1 was dissolved in THF(406 mL), and lithium hydroxide hydrate (51.9 g), water (309 mL) andmethanol (77 mL) were added thereto, and the mixture was stirred at roomtemperature for 3 hr. Ice block was added to the reaction solution, andthen 5N hydrochloric acid (263 mL) was added thereto. Sodium chlorideand 10% methanol/dichloromethane solution were added thereto, and themixture was subjected to extraction. 10% Methanol/dichloromethanesolution was added to the aqueous layer, and the mixture was subjectedto extraction. The combined organic layers were dried over anhydroussodium sulfate, and the solvent was evaporated under reduced pressure togive the title compound (36.8 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.38 (9H, s), 1.46-1.56 (1H, m), 1.60-1.69 (1H, m),1.90-1.99 (1H, m), 2.08-2.20 (1H, m), 2.76-2.86 (1H, m), 3.48-3.58 (1H,m), 3.77-3.84 (1H, m), 4.80 (1H, d, J=4.5 Hz), 6.76 (1H, d, J=7.5 Hz),12.10 (1H, br s).

Step 3 methoxymethyl(1R,3S,4S)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylate

The title compound was obtained in the same manner as in Step 8 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.44 (9H, s), 1.69-1.79 (1H, m), 2.06-2.17 (2H, m),2.39-2.51 (1H, m), 3.02-3.12 (1H, m), 3.37 (3H, s), 3.48 (3H, s),3.93-4.06 (2H, m), 4.64 (1H, d, J=7.0 Hz), 4.72 (1H, d, J=7.0 Hz), 5.05(1H, br s), 5.25 (2H, s).

MS (m/z): 234 (M-Boc+H)⁺.

Step 4(1R,3S,4S)-3-[(tert-butoxycarbonyl)amino]-4-(methoxymethoxy)cyclopentane-1-carboxylicacid

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound obtained in the above Step 3.

Step 5 benzyl tert-butyl[(1R,3S,4S)-4-(methoxymethoxy)cyclopentane-1,3-diyl]biscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using the compound obtained in the above Step 4.

¹H-NMR (CDCl₃) δ: 1.36-1.51 (1H, m), 1.44 (9H, s), 1.91-2.04 (2H, m),2.51-2.61 (1H, m), 3.35 (3H, s), 3.77 (1H, br s), 4.03-4.13 (2H, m),4.62 (1H, d, J=6.7 Hz), 4.66-4.72 (1H, m), 5.15-5.22 (1H, m), 5.09 (2H,s), 7.32-7.36 (5H, m).

MS (m/z): 295 (M-Boc+H)⁺.

Step 6 benzyl [(1R,3S,4S)-3-amino-4-hydroxycyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 4 ofReference Example A-3, using the compound obtained in the above Step 5.

¹H-NMR (CDCl₃) δ: 1.25-1.33 (1H, m), 2.02 (2H, t, J=6.1 Hz), 2.34-2.42(1H, m), 3.19-3.24 (1H, m), 3.95 (1H, dt, J=4.9, 4.9 Hz), 4.22-4.30 (1H,m), 5.08 (2H, s), 5.36 (1H, br s), 7.30-7.40 (5H, m).

MS (m/z): 251 (M+H)⁺.

Step 7 benzyl{(1R,3S,4S)-3-hydroxy-4-[(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 12 ofReference Example A-1, using the compound obtained in the above Step 6.

¹H-NMR (CDCl₃) δ: 1.45-1.53 (1H, m), 2.01 (2H, t, J=7.1 Hz), 2.35 (1H,br s), 2.41-2.51 (1H, m), 3.49 (1H, br s), 4.07 (1H, dd, J=15.0, 7.7Hz), 4.26 (1H, br s), 4.86 (1H, br s), 5.08 (2H, s), 6.02 (1H, br s),7.30-7.39 (5H, m), 7.74-7.78 (2H, m), 7.87 (1H, dd, J=6.1, 3.1 Hz), 8.15(1H, dd, J=5.5, 3.1 Hz).

MS (m/z): 436 (M+H)⁺.

Step 8 benzyl{(1R,3S,4S)-3-hydroxy-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1, using the compound obtained in the above Step 7.

¹H-NMR (CDCl₃) δ: 1.83-1.92 (1H, m), 2.00-2.08 (1H, m), 2.22-2.30 (1H,m), 2.46 (1H, br s), 2.90 (3H, s), 3.94 (1H, dt, J=11.7, 7.4 Hz),4.05-4.11 (1H, m), 4.27 (1H, br s), 4.76 (1H, br s), 5.07 (2H, s),7.29-7.38 (5H, m), 7.63-7.67 (1H, m), 7.69-7.74 (2H, m), 8.07-8.10 (1H,m).

MS (m/z): 450 (M+H)⁺.

Step 9 benzyl [(1R,3S,4S)-3-hydroxy-4-(methylamino)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 14 ofReference Example A-1, using the compound obtained in the above Step 8.

¹H-NMR (CDCl₃) δ: 1.34 (1H, dt, J=13.5, 5.5 Hz), 1.96-2.01 (2H, m),2.32-2.39 (1H, m), 2.41 (3H, s), 2.85-2.90 (1H, m), 4.05-4.09 (1H, m),4.27 (1H, br s), 5.08 (2H, s), 5.39 (1H, br s), 7.30-7.39 (5H, m).

MS (m/z): 265 (M+H)⁺.

Reference Example A-15 tert-butyl[(1R,3R,4S)-3-hydroxy-4-(methylamino)cyclopentyl]carbamate Step 1tert-butyl{(1R,3R,4S)-3-hydroxy-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The compound (20.6 g) obtained in Step 13 of Reference Example A-1 wasdissolved in acetonitrile (305 mL), and iodotrimethylsilane (18.8 mL)was added dropwise thereto under ice-cooling, and the mixture wasstirred at the same temperature for 1 hr. 1N Hydrochloric acid ethanolsolution (45.8 mL) was added to the reaction solution, and the mixturewas stirred at the same temperature for 30 min, and concentrated underreduced pressure. The residue was dissolved in ethanol (15 mL), and THF(153 mL), sodium carbonate (24.3 g) and di-tert-butyl dicarbonate (15.0g) were added thereto, and the mixture was stirred at room temperaturefor 3 hr. Ethyl acetate and saturated brine were added to the reactionsolution, and the mixture was subjected to extraction operation. Theorganic layer was dried over anhydrous sodium sulfate, and the solventwas evaporated under reduced pressure. The residue was purified bysilica gel chromatography (n-hexane/ethyl acetate) to give the titlecompound (17.1 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.45 (9H, s), 1.57-1.65 (1H, m), 1.97-2.20 (2H, m),2.23-2.32 (1H, m), 3.08 (3H, s), 3.28-3.32 (1H, m), 3.84-3.94 (2H, m),4.20-4.26 (1H, m), 5.00 (1H, br s), 7.63-7.67 (1H, m), 7.67-7.75 (2H,m), 8.02-8.05 (1H, m).

Step 2 tert-butyl[(1R,3R,4S)-3-hydroxy-4-(methylamino)cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 14 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.32 (1H, ddd, J=13.0, 9.5, 5.5 Hz), 1.42 (9H, s),1.80 (1H, d, J=14.5 Hz), 1.99 (1H, ddd, J=14.5, 8.5, 4.0 Hz), 2.34-2.44(4H, m), 2.81 (1H. td, J=8.5, 4.0 Hz), 3.99 (1H, td, J=4.0, 2.0 Hz),4.04-4.15 (1H, m), 5.19-5.31 (1H, m).

Reference Example A-16

tert-butyl [(1S,2R,4R)-4-amino-2-methoxycyclopentyl]methylcarbamate

Step 1 methyl(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-methoxycyclopentane-1-carboxylate

A mixture of methyl(1R,3S,4R)-3-(tert-butoxycarbonylamino)-4-hydroxy-cyclopentanecarboxylate(CAS: 321744-23-4) (3.55 g), dichloromethane (60 mL), silver(I) oxide(CAS: 20667-12-3) (9.18 g), molecular sieves 3A (CAS: 308080-99-1) (4.73g, used after dried under reduced pressure at 180° C. for 2 hr) andmethyl iodide (CAS: 74-88-4) (16.2 mL) was stirred vigorously at 55° C.for 42 hr. The reaction solution was filtered through Celite, and thefiltrate was concentrated under reduced pressure. The obtained residuewas purified by silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (3.25 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.45 (9H, s), 1.85 (1H, dt, J=16.8, 6.4 Hz), 1.93-2.03(1H, m), 2.15-2.23 (1H, m), 2.29 (1H, dt, J=14.9, 6.4 Hz), 2.75-2.84(1H, m), 3.30 (3H, s), 3.59-3.66 (1H, m), 3.68 (3H, s), 3.88-3.98 (1H,m), 5.12 (1H, d, J=7.4 Hz).

Step 2(1R,3S,4R)-3-[(tert-butoxycarbonyl)amino]-4-methoxycyclopentane-1-carboxylicacid

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.45 (9H, s), 1.83-1.94 (1H, m), 1.94-2.03 (1H, m),2.23 (1H, ddd, J=14.7, 5.0, 2.4 Hz), 2.28-2.39 (1H, m), 2.81-2.91 (1H,m), 3.32 (3H, s), 3.65 (1H, td, J=4.9, 2.4 Hz), 3.89-4.01 (1H, m), 5.12(1H, d, J=6.1 Hz).

Step 3 benzyl tert-butyl[(1R,3S,4R)-4-methoxycyclopentane-1,3-diyl]biscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.34-1.43 (1H, m), 1.45 (9H, s), 1.73-1.83 (1H, m),1.94-2.04 (1H, m), 2.48-2.60 (1H, m), 3.32 (3H, s), 3.60-3.65 (1H, m),3.84-3.96 (1H, m), 4.12-4.20 (1H, m), 5.00-5.16 (4H, m), 7.30-7.38 (5H,m).

Step 4 benzyl{(1R,3R,4S)-3-methoxy-4-[(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 11 andStep 12 of Reference Example A-1, using the compound obtained in theabove Step 3.

¹H-NMR (CDCl₃) δ: 1.43-1.52 (1H, m), 1.69-1.76 (1H, m), 1.94-2.03 (1H,m), 2.35 (1H, dt, J=15.3, 6.9 Hz), 3.14 (3H, s), 3.45-3.50 (1H, m),3.73-3.81 (1H, m), 4.09-4.19 (1H, m), 5.01 (1H, d, J=8.6 Hz), 5.05 (2H,s), 6.04 (1H, d, J=8.6 Hz), 7.29-7.38 (5H, m), 7.71-7.78 (2H, m),7.87-7.91 (1H, m), 8.13-8.18 (1H, m).

MS (m/z):450 (M+H)⁺.

Step 5 benzyl{(1R,3R,4S)-3-methoxy-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1, using the compound obtained in the above Step 4.

¹H-NMR (CDCl₃) δ: 1.66 (1H, dd, J=14.7, 2.5 Hz), 1.76 (1H, td, J=12.1,6.3 Hz), 1.99-2.10 (1H, m), 2.22-2.34 (1H, m), 3.01 (3H, s), 3.29 (3H,s), 3.74-3.80 (1H, m), 4.02-4.24 (2H, m), 5.03-5.16 (3H, m), 7.29-7.40(5H, m), 7.60-7.73 (3H, m), 7.96-8.03 (1H, m).

MS (m/z):464 (M+H)⁺.

Step 6 benzyl [(1R,3R,4S)-3-methoxy-4-(methylamino)cyclopentyl]carbamate

The title compound was obtained as a crude product in the same manner asin Step 14 of Reference Example A-1, using the compound obtained in theabove Step 5.

Step 7 benzyl{(1R,3S,4R)-3-[(tert-butoxycarbonyl)(methyl)amino]-4-methoxycyclopentyl}carbamate

The compound (4.82 g) obtained in the above Step 6 was dissolved in THF(40 mL), and a solution of di-tert-butyl dicarbonate (CAS: 24424-99-5)(3.02 g) in THF (20 mL), and TEA (CAS: 121-44-8) (2.3 mL) were addedthereto, and the mixture was stirred at room temperature for 16 hr. Thesolvent was evaporated under reduced pressure, and the obtained residuewas purified by silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (3.15 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.47 (9H, s), 1.59-1.66 (1H, m), 1.80 (1H, td, J=12.1,7.0 Hz), 2.02-2.14 (1H, m), 2.25 (1H, dt, J=14.9, 6.4 Hz), 2.89 (3H, s),3.27 (3H, s), 3.71-3.80 (1H, m), 4.01-4.32 (2H, m), 5.05-5.18 (3H, m),7.30-7.38 (5H, m).

Step 8 tert-butyl[(1S,2R,4R)-4-amino-2-methoxycyclopentyl]methylcarbamate

A mixture of the compound (3.15 g) obtained in the above Step 7, 5%palladium on carbon (PH) wet (CAS: 7440-05-3) (3.41 g) and ethanol (100mL) was stirred under hydrogen atmosphere at room temperature for 3.5hr. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure. The obtained residue was purifiedby amino silica gel column chromatography (ethyl acetate/methanol) togive the title compound (1.78 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.40-1.47 (1H, m), 1.47 (9H, s), 1.68-1.79 (1H, m),2.07 (1H, dt, J=13.3, 6.1 Hz), 2.12-2.22 (1H, m), 2.92 (3H, s),3.21-3.31 (1H, m), 3.28 (3H, s), 3.69-3.81 (1H, m), 3.95-4.34 (1H, m).Reference Example A-17

tert-butyl [(1S,2S,4R)-4-amino-2-methoxycyclopentyl]methylcarbamate

Step 1 benzyl{(1R,3S,4S)-3-[(tert-butoxycarbonyl)(methyl)amino]-4-hydroxycyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 7 ofReference Example A-16, using the compound obtained in Step 9 ofReference Example A-14.

¹H-NMR (CDCl₃) δ: 1.47 (9H, s), 1.57-1.69 (1H, m), 1.82-1.94 (1H, m),1.95-2.06 (1H, m), 2.27-2.40 (1H, m), 2.83 (3H, s), 3.89-4.01 (1H, m),4.09-4.21 (1H, m), 4.22-4.33 (1H, m), 5.09 (3H, br s), 7.29-7.38 (5H,m).

Step 2 benzyl{(1R,3S,4S)-3-[(tert-butoxycarbonyl)(methyl)amino]-4-methoxycyclopentyl}carbamate

The compound (1.10 g) obtained in the above Step 1 was dissolved in THF(37 mL), and methyl iodide (0.31 mL) and sodium hydride (CAS: 7646-69-7)(purity 55%, 0.205g) were added thereto, and the mixture was stirred at0° C. for 1 hr. Then, additional methyl iodide (0.10 mL) and sodiumhydride (purity 55%, 0.067g) were added thereto, and the mixture wasstirred at 0° C. for 1 hr. Water was added to the reaction solution, andthe mixture was extracted with ethyl acetate. The organic layer wasdried over anhydrous sodium sulfate, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(n-hexane/acetone) to give the title compound (0.827 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.46 (9H, s), 1.55-1.71 (1H, m), 1.76-2.04 (2H, m),2.25-2.39 (1H, m), 2.86 (3H, br s), 3.30 (3H, s), 3.71-4.15 (3H, m),5.10 (2H, s), 7.29-7.38 (5H, m).

Step 3 tert-butyl[(1S,2S,4R)-4-amino-2-methoxycyclopentyl]methylcarbamate

The title compound was obtained in the same manner as in Step 8 ofReference Example A-16, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.34-1.45 (1H, m), 1.47 (9H, s), 1.64 (1H, dt, J=15.1,6.7 Hz), 1.89-1.97 (1H, m), 2.10-2.19 (1H, m), 2.83 (3H, s), 3.30 (3H,s), 3.37-3.47 (1H, m), 3.87 (1H, dt, J=8.8, 4.0 Hz), 4.17-4.31 (1H, m).

Reference Example A-18 tert-butyl[(1R,5S)-5-amino-3,3-difluorocyclohexyl]carbamate Step 1 benzyltert-butyl [(1R,3S)-5,5-difluorocyclohexane-1,3-diyl]biscarbamate

The title compound was obtained in the same manner as in Step 10 ofReference Example A-1, using(1S,5R)-5-[(tert-butoxycarbonyl)amino]-3,3-difluorocyclohexane-1-carboxylicacid (CAS: 2227198-19-6).

¹H-NMR (DMSO-D₆) δ: 1.16-1.28 (1H, m), 1.38 (9H, s), 1.58-1.69 (2H, m),1.93-1.96 (1H, m), 2.19-2.20 (2H, m), 3.48-3.49 (2H, m), 5.02 (2H, s),7.10-7.12 (1H, m), 7.20-7.22 (1H, m), 7.32-7.37 (4H, m), 7.48-7.50 (1H,m).

MS (m/z): 285 (M-Boc+H)⁺.

Step 2 tert-butyl [(1R,5S)-5-amino-3,3-difluorocyclohexyl]carbamate

To a solution of the compound (596 mg) obtained in the above Step 1 inethanol (20 mL) was added 10% palladium on carbon catalyst wet (400 mg),and the mixture was stirred under hydrogen atmosphere at roomtemperature for 4 hr. After nitrogen substitution, the mixture wasfiltered through Celite, and the filtrate was concentrated under reducedpressure to give the title compound (445 mg) as an oil. The obtainedcompound was directly used in the next step.

MS (m/z): 251 (M+H)⁺.

Reference Example A-19 tert-butyl{(1S,3R)-3-[(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)amino]cyclopentyl}carbamateStep 1 tert-butyl{(1S,3R)-3-[(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1 except that(2-bromoethoxy)-tert-butyldimethylsilane (CAS: 86864-60-0) was usedinstead of iodomethane, using the compound obtained in Step 2 ofReference

Example A-3

¹H-NMR (DMSO-D₆) δ: 0.05 (6H, s), 0.87 (9H, s), 1.35 (9H, s), 1.51-1.80(6H, m), 3.31-3.33 (2H, m), 3.68-3.70 (3H, m), 4.03-4.06 (1H, m), 6.94(1H, s), 7.82-8.08 (4H, m).

MS (m/z): 444 (M-Boc+H)⁺.

Step 2 tert-butyl{(1S,3R)-3-[(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)amino]cyclopentyl}carbamate

A mixture of the compound (585 mg) obtained in the above Step 1,4-tert-butylbenzenethiol (0.362 mL), potassium carbonate (595 mg) andDMF (5.4 mL) was stirred at 40° C. 6 hr. Water was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate) togive the title compound (428 mg) as an oil.

¹H-NMR (DMSO-D₆) δ: 0.04 (6H, s), 0.87 (9H, s), 1.16-1.23 (1H, m), 1.37(9H, s), 1.43-1.47 (2H, m), 1.72-1.73 (2H, m), 2.00-2.03 (1H, m), 2.60(2H, t, J=5.8 Hz), 3.01-3.03 (1H, m), 3.62 (2H, t, J=5.8 Hz), 3.71-3.73(1H, m), 6.79 (1H, d, J=7.7 Hz).

MS (m/z): 359 (M+H)⁺.

Reference Example B-14-chloro-2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine Step 12-amino-5-(2,2,2-trifluoroethyl)thiophene-3-carboxamide

To 4,4,4-trifluorobutanal hydrate (2.00 g), 2-cyanoacetamide (CAS:107-91-5) (1.75 g) and sulfur (668 mg) was added DMF (14 mL), and TEA(3.46 mL) was added dropwise thereto under ice-cooling. After thecompletion of dropwise addition, the mixture was allowed to warm to roomtemperature, stirred for 10 hr, and allowed to stand overnight. Ethylacetate and saturated brine were added to the reaction solution, and themixture was subjected to liquid separation. The organic layer was washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (3.00 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.59 (2H, q, J=11.1 Hz), 6.75 (1H, br s), 7.00 (1H,s), 7.21 (1H, br s), 7.30 (2H, s).

MS (m/z): 225 (M+H)⁺.

Step 2 2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4(3H)-one

To the compound (2.95 g) obtained in the above Step 1 were added aceticacid (13 mL) and triethyl orthoacetate (CAS: 78-39-7) (10 mL), and themixture was heated under reflux for 3 hr. Then, the mixture was heatedin a microwave reactor (at 150° C. for 5.5 hr). Saturated aqueous sodiumhydrogencarbonate solution and ethyl acetate were added to the reactionsolution, and the mixture was subjected to liquid separation. Theaqueous layer was extracted three times with ethyl acetate, and theorganic layer was dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (n-hexane/ethyl acetate, dichloromethane/methanol)to give the title compound (0.86 g) as a solid.

¹H-NMR (CDCl₃) δ: 2.59 (3H, s), 3.62 (2H, q, J=10.1 Hz), 7.39 (1H, s),12.35 (1H, br s).

Step 3 4-chloro-2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine

To 2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4(3H)-one(0.86 g) were added phosphorus oxychloride (10.33 g) and DMF (0.03 mL),and the mixture was stirred at 110° C. for 3.5 hr. The reaction solutionwas added little by little to a mixture of dichloromethane and ice, andthe mixture was stirred vigorously for 1 hr, and subjected to liquidseparation. The organic layer was washed with water, dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (0.89 g) as an oil.

¹H-NMR (CDCl₃) δ: 2.81 (3H, s), 3.71 (2H, q, J=10.0 Hz), 7.32 (1H, s).

Reference Example B-24-chloro-6-(cyclopropylmethyl)thieno[2,3-d]pyrimidine Step 16-(cyclopropylmethyl)thieno[2,3-d]pyrimidin-4(3H)-one

To 3-cyclopropylpropanal (CAS: 5618-02-0) (968 mg), 2-cyanoacetamide(829 mg) and sulfur (316 mg) was added DMF (10 mL), and the mixture wascooled well in an ice-salt bath. TEA (1.64 mL) was added dropwisethereto. After the completion of dropwise addition, and the mixture wasallowed to warm to room temperature, and stirred for 10 hr. The solventwas evaporated under reduced pressure. Acetic acid (10 mL) and triethylorthoformate (6.8 mL) were added to the residue, and the mixture washeated under reflux for 3.5 hr. The solvent was evaporated under reducedpressure, and a mixed solvent of n-hexane/ethyl acetate=1/1 was added tothe residue. The solid was collected by filtration, and washed with amixed solvent of n-hexane/ethyl acetate=1/1 to give the title compound(1.32 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 0.24-0.29 (2H, m), 0.51-0.57 (2H, m), 0.99-1.08 (1H,m), 2.75 (2H, d, J=6.7 Hz), 7.16 (1H, s), 8.06 (1H, s), 12.43 (1H, brs).

MS (m/z): 207 (M+H)⁺.

Step 2 4-chloro-6-(cyclopropylmethyl)thieno[2,3-d]pyrimidine

The title compound was obtained in the same manner as in Step 3 ofReference Example B-1, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 0.32-0.36 (2H, m), 0.67-0.71 (2H, m), 1.08-1.18 (1H,m), 2.86 (2H, d, J=7.3 Hz), 7.18 (1H, s), 8.78 (1H, s).

MS (m/z): 225, 227 (M+H)⁺.

Reference Example B-3 4-chloro-6-cyclopropylthieno[2,3-d]pyrimidine

A mixture of 6-bromo-4-chlorothieno[2,3-d]pyrimidine (100 mg),cyclopropylboronic acid (79.0 mg), sodium carbonate (144 mg), toluene(1.5 mL), water (0.5 mL) and tetrakis(triphenylphosphine)palladium(0)(56.0 mg) was stirred under nitrogen atmosphere at 110° C. Water wasadded to the reaction solution, and the mixture was extracted withdichloromethane. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (36.8 mg) as asolid.

¹H-NMR (CDCl₃) δ: 0.92-0.96 (2H, m), 1.20-1.23 (2H, m), 2.19-2.26 (1H,m), 7.05 (1H, s), 8.75 (1H, s).

MS (m/z): 211, 213 (M+H)+.

Reference Example B-42,4-dichloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine Step 16-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine-2,4(1H,3H)-dione

The compound (6.20 g) obtained in Step 1 of Reference Example B-1 wasdissolved in 1,4-dioxane (100 mL), and triphosgene (3.55 g) was addedthereto, and the mixture was heated under reflux for 6 hr. The reactionsolution was allowed to cool to room temperature, and concentrated underreduced pressure. The residue was subjected to slurry washing withdichloromethane to give the title compound (2.41 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.93 (2H, q, J=11.0 Hz), 7.14 (1H, s), 11.19 (1H,s), 11.92 (1H, s).

Step 2 2,4-dichloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine

The compound (2.41 g) obtained in the above Step 1 was suspended inphosphorus oxychloride (11.5 mL), and DMF (0.030 mL) was added thereto,and the mixture was stirred with heating at 110° C. for 4 hr. Thereaction solution was allowed to cool to room temperature, washed withdichloromethane, and poured into ice water, and the mixture was stirredvigorously at room temperature for 1 hr. The organic layer was driedover anhydrous sodium sulfate, and the solvent was evaporated underreduced pressure. The residue was purified by silica gel chromatography(dichloromethane/n-hexane) to give the title compound (1.42 g) as asolid.

¹H-NMR (CDCl₃) δ: 3.74 (2H, q, J=10.0 Hz), 7.37 (1H, s).

Reference Example B-54-chloro-2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine Step12-(methylsulfanyl)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4(3H)-one

A mixture of the compound (40.0 g) obtained in Step 1 of ReferenceExample B-1, ethanol (500 mL) and potassium ethylxanthate (85.8 g) washeated under reflux under nitrogen atmosphere for 19 hr. The reactionsolution was concentrated under reduced pressure, and the residue wassubjected to slurry washing with dichloromethane (200 mL), and the solid(101 g) was collected by filtration. To a mixture of the obtained solid(101 g) and DMF (2.09 L) was added methyl iodide (13.0 mL) underice-cooling, and the mixture was stirred at room temperature for 1 hr.Methyl iodide (8.5 mL) was added thereto, and the mixture was stirredfor additional 30 min. Water was added to the reaction solution, and theprecipitated solid was collected by filtration to give the titlecompound (14.5 g) as a solid. Ethyl acetate was added to the filtrate,and the mixture was subjected to extraction. The organic layer waswashed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. Water was added thereto, and theprecipitated solid was collected by filtration to give the titlecompound (3.85 g) as a solid. The filtrate was acidified withhydrochloric acid, and extracted with ethyl acetate. The organic layerwas washed with saturated brine, dried over anhydrous sodium sulfate,and concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (17.1 g) as a solid.

¹H-NMR (CDCl₃) δ: 2.65 (3H, s), 3.59 (2H, q, J=10.0 Hz), 7.36 (1H, s),11.35 (1H, s).

MS (m/z): 281 (M+H)⁺.

Step 22-(methanesulfonyl)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4(3H)-one

To a mixture of the compound (42.2 g) obtained in the above Step 1 andTHF (600 mL) was added a mixture of Oxone (CAS: 10058-23-8) (278 g) andwater (600 mL) under ice-cooling, and the mixture was stirred at roomtemperature for 4 hr. Water was added to the reaction solution, and theprecipitated solid was collected by filtration to give the titlecompound (43.7 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.44 (3H, s), 4.20 (2H, q, J=11.0 Hz), 7.56 (1H, s).

MS (m/z): 313 (M+H)⁺.

Step 32-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4(1H)-one

To the compound (14.3 g) obtained in the above Step 2 were addedmethanol (450 mL) and potassium carbonate (12.7 g), and the mixture washeated under reflux for 2 hr. Additional potassium carbonate (6.33 g)was added thereto, and the mixture was heated under reflux for 1 hr. Thereaction solution was concentrated to about one-half to one-thirdvolume, and acidified with 2N hydrochloric acid. Ethyl acetate was addedthereto, and the mixture was subjected to liquid separation, and theaqueous layer was extracted with ethyl acetate. The organic layers werecombined, washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. Diethyl ether wasadded to the obtained residue, and the solid was collected by filtrationto give the title compound (7.85 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.57 (2H, q, J=10.3 Hz), 4.06 (3H, s), 7.33 (1H, s),11.08 (1H, br s).

Step 44-chloro-2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine

To a suspension of the compound (8.39 g) obtained in the above Step 3 inphosphorus oxychloride (36.9 mL) was added DMF (8 drops), and themixture was stirred at room temperature for 2.5 hr, and then at 60° C.for 15 min. The reaction solution was allowed to cool, and added littleby little to a mixture of sodium hydrogencarbonate and ice water, andthe used container was washed with dichloromethane. The mixture wasstirred for 10 min, dichloromethane was added thereto, and the mixturewas subjected to liquid separation. The aqueous layer was adjusted topH=ca.9 with saturated aqueous sodium bicarbonate solution, andextracted with dichloromethane. The combined organic layers were washedwith saturated aqueous sodium bicarbonate solution, dried over sodiumsulfate, and concentrated under reduced pressure. The obtained residuewas purified by silica gel column chromatography (ethylacetate/n-hexane) to give the title compound (7.85 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.66 (2H, q, J=10.1 Hz), 4.09 (3H, s), 7.24 (1H, s).

MS (m/z): 283, 285 (M+H)⁺.

Reference Example B-64-chloro-6-(2,2,2-trifluoroethyl)thieno[3,2-d]pyrimidine Step 14-chloro-6-{2,2,2-trifluoro-1-[(trimethylsilyl)oxy]ethyl}thieno[3,2-d]pyrimidine

4-Chlorothieno[3,2-d]pyrimidine-6-carbaldehyde (CAS: 875340-14-0) (500mg) was dissolved in THF (6 mL), and (trifluoromethyl)trimethylsilane(CAS: 81290-20-2) (0.558 mL) was added thereto, and thentetrabutylammonium fluoride (1.0 mol/L, THF solution) (0.126 mL) wasadded thereto under ice-cooling. The mixture was allowed to cool to roomtemperature, and stirred for 1 hr. Ethyl acetate and saturated brinewere added to the reaction solution, and the mixture was subjected toliquid separation. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure. The obtained residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (294mg) as an oil.

¹H-NMR (CDCl₃) δ: 0.25 (9H, s), 5.37 (1H, q, J=6.0 Hz), 7.60 (1H, s),9.00 (1H, s).

MS (m/z): 341, 343 (M+H)⁺.

Step 2 1-(4-chlorothieno[3,2-d]pyrimidin-6-yl)-2,2,2-trifluoroethan-1-ol

The compound (260 mg) obtained in the above Step 1 was dissolved in THF(15 mL), 1N hydrochloric acid (1.1 mL) was added thereto, and themixture was stirred at room temperature for 15 min. Ethyl acetate andsaturated aqueous sodium hydrogencarbonate solution were added to thereaction solution, and the mixture was subjected to liquid separation.The organic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure to give thetitle compound (206 mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.43 (1H, s), 5.50-5.55 (1H, m), 7.68 (1H, s), 9.01(1H, s).

MS (m/z): 269, 271 (M+H)⁺.

Step 3 O-[1-(4-chlorothieno[3,2-d]pyrimidin-6-yl)-2,2,2-trifluoroethyl]O-phenyl carbonothioate

To a dichloromethane solution (1 mL) of phenyl chlorothionocarbonate(CAS: 1005-56-7) (77 mg) were added the compound (100 mg) obtained inthe above Step 2 and TEA (0.067 mL), and the mixture was stirred at roomtemperature for 2 hr. Water and dichloromethane were added to thereaction solution, and the mixture was subjected to liquid separation.The organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (125 mg) as a solid.

¹H-NMR (CDCl₃) δ: 7.07-7.15 (3H, m), 7.33 (1H, t, J=7.6 Hz), 7.44 (2H,t, J=7.6 Hz), 7.84 (1H, s), 9.05 (1H, s).

MS (m/z): 405, 407 (M+H)⁺.

Step 4 4-chloro-6-(2,2,2-trifluoroethyl)thieno[3,2-d]pyrimidine

The compound (2.42 g) obtained in the above Step 3 was dissolved intoluene (120 mL), tributyltin hydride (CAS: 688-73-3) (3.45 mL) and2,2′-azobis(isobutyronitrile) (CAS: 78-67-1) (393 mg) were addedthereto, and the mixture was stirred at 80° C. for 2 hr. The solvent wasevaporated under reduced pressure, and the residue was purified bysilica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (1.22 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.81 (2H, q, J=10.1 Hz), 7.55 (1H, s), 9.00 (1H, s).

MS (m/z): 253, 255 (M+H)⁺.

Reference Example B-7 6-(2,2,2-trifluoroethyl)quinazolin-4(3H)-one Step1 (4-methoxyquinazolin-6-yl)methanol

A mixture of methyl 4-chloroquinazoline-6-carboxylate (CAS: 152536-17-9)(3.00 g) and THF (30 mL) was cooled to −50° C., diisobutylaluminiumhydride (CAS: 1191-15-7) (1.0 mol/L, toluene solution, 29.0 mL) wasadded slowly thereto, and the mixture was warmed to 0° C. over 1.5 hr.Then, the mixture was allowed to warm to room temperature, and stirredfor 20 hr. Then, the reaction solution was cooled to 0° C.,diisobutylaluminium hydride (1.0 mol/L, toluene solution, 15.0 mL) wasadded slowly thereto, and the mixture was stirred at the sametemperature for 15 min, and then at room temperature for 2 hr. Thereaction solution was cooled again to 0° C., aqueous potassium sodiumtartrate solution (2 mol/L, 75 mL) was added thereto. The mixture wasallowed to warm to room temperature, and stirred overnight. The reactionmixture was extracted with ethyl acetate and dichloromethane, and theorganic layer was dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was subjected to silica gel columnchromatography (n-hexane/ethyl acetate). The obtained residue wasdissolved in methanol, and the solution was concentrated under reducedpressure to give the title compound (1.46 g, containing impurities(purity ca.60%)) as a solid. This compound was used in the next reactionwithout further purification.

¹H-NMR (DMSO-D₆) δ: 4.20 (3H, s), 4.72 (2H, s), 7.96 (2H, s), 8.14 (1H,s), 8.96 (1H, s).

MS (m/z): 191 (M+H)⁺.

Step 2 4-methoxyquinazoline-6-carbaldehyde

A mixture of the compound (1.45 g, containing impurities (purityca.60%)) obtained in the above Step 1, dichloromethane (60 mL) andDess-Martin periodinane (CAS: 87413-09-0) (3.51 g) was stirred at roomtemperature for 3 hr. Saturated aqueous sodium bicarbonate solution wasadded to the reaction solution, and the mixture was extracted withdichloromethane. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure, and the residue was subjected to silica gel columnchromatography (n-hexane/ethyl acetate). The obtained solid wasdissolved in ethyl acetate, and the solution was washed with saturatedaqueous sodium bicarbonate solution, water and saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressureto give the title compound (0.893 g, containing impurities (purityca.60%)) as a solid. This compound was used in the next reaction withoutfurther purification.

¹H-NMR (CDCl₃) δ: 4.25 (3H, s), 8.05 (1H, d, J=8.5 Hz), 8.33 (1H, dd,J=8.5, 1.8 Hz), 8.70 (1H, d, J=1.8 Hz), 8.93 (1H, s), 10.18 (1H, s).

MS (m/z): 189 (M+H)⁺.

Step 34-methoxy-6-{2,2,2-trifluoro-1-[(trimethylsilyl)oxy]ethyl}quinazoline

To a mixture of the compound (0.890 g, containing impurities (purityca.60%)) obtained in the above Step 2 and THF (23 mL) were added(trifluoromethyl)trimethylsilane (1.1 mL) and cesium fluoride (CAS:13400-13-0) (0.0336 g) under ice-cooling. The mixture was stirred at thesame temperature for 10 min, and then at room temperature for 5 hr.Then, additional cesium fluoride (0.240 g) was added thereto, and themixture was stirred at room temperature for 3.5 hr. Then, additionalcesium fluoride (0.240 g) was added thereto, and the mixture was stirredat room temperature for 14 hr. Then, additional(trifluoromethyl)trimethylsilane (0.37 mL) and cesium fluoride (0.250 g)were added thereto, and the mixture was stirred at room temperature for5 hr. Saturated brine was added to the reaction solution, and themixture was extracted with ethyl acetate. The organic layer was washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (0.633 g) as a solid.

¹H-NMR (CDCl₃) δ: 0.15 (9H, s), 4.21 (3H, s), 5.09 (1H, q, J=6.5 Hz),7.93-7.99 (2H, m), 8.23 (1H, s), 8.84 (1H, s).

Step 4 2,2,2-trifluoro-1-(4-methoxyquinazolin-6-yl)ethan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example B-6, using the compound obtained in the above Step 3.

¹H-NMR (DMSO-D₆) δ: 4.16 (3H, s), 5.46-5.56 (1H, m), 7.16 (1H, d, J=6.1Hz), 7.97 (1H, d, J=8.5 Hz), 8.05 (1H, dd, J=8.5, 1.2 Hz), 8.32 (1H, d,J=1.2 Hz), 8.85 (1H, s).

Step 5 O-phenyl O-[2,2,2-trifluoro-1-(4-methoxyquinazolin-6-yl)ethyl]carbonothioate

The title compound was obtained in the same manner as in Step 3 ofReference Example B-6, using the compound obtained in the above Step 4.

¹H-NMR (CDCl₃) δ: 4.23 (3H, s), 6.77 (1H, q, J=6.7 Hz), 7.09-7.13 (2H,m), 7.29-7.34 (1H, m), 7.39-7.46 (2H, m), 7.98 (1H, dd, J=8.5, 1.8 Hz),8.04 (1H, d, J=8.5 Hz), 8.37 (1H, d, J=1.8 Hz), 8.87 (1H, s).

MS (m/z): 395 (M+H)⁺.

Step 6 4-methoxy-6-(2,2,2-trifluoroethyl)quinazoline

The title compound was obtained in the same manner as in Step 4 ofReference Example B-6, using the compound obtained in the above Step 5.

1H-NMR (CDCl₃) δ: 3.56 (2H, q, J=10.7 Hz), 4.20 (3H, s), 7.77 (1H, dd,J=8.5, 1.8 Hz), 7.95 (1H, d, J=8.5 Hz), 8.12 (1H, d, J=1.8 Hz), 8.83(1H, s).

Step 7 6-(2,2,2-trifluoroethyl)quinazolin-4(3H)-one

The compound (0.111 g) obtained in the above Step 6 was dissolved in THF(2.8 mL), and the solution was added 1N hydrochloric acid (1.4 mL) underice-cooling. The reaction solution was warmed to room temperature over5.5 hr, and saturated aqueous sodium bicarbonate solution was addedthereto. The reaction mixture was extracted with dichloromethane. Theobtained aqueous layer was acidified with 1N hydrochloric acid, andextracted with dichloromethane. All of the organic layers were combined,dried over anhydrous sodium sulfate, and concentrated under reducedpressure to give the title compound (0.101 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.86 (2H, q, J=11.5 Hz), 7.69 (1H, d, J=8.5 Hz),7.79 (1H, dd, J=8.5, 1.8 Hz), 8.12 (1H, s), 8.14 (1H, d, J=1.8 Hz),12.32 (1H, s).

MS: m/z 229 (M+H)⁺.

Reference Example B-8 6-(methoxymethyl)thieno[2,3-d]pyrimidin-4(3H)-one

The title compound was obtained in the same manner as in Step 1 ofReference Example B-2 except that 3-methoxypropanal (1.00 g)(CAS2806-84-0) was used instead of 3-cyclopropylpropanal.

¹H-NMR (DMSO-D₆) δ: 3.30 (3H, s), 4.63 (2H, s), 7.33 (1H, s), 8.12 (1H,s), 11.42 (1H, br s).

MS (m/z): 197 (M+H)⁺.

Reference Example B-9 6-(oxetan-3-yl)thieno[2,3-d]pyrimidin-4(3H)-one

The title compound was obtained in the same manner as in Step 1 ofReference Example B-2 except that (oxetan-3-yl)acetaldehyde synthesizedaccording to the method described in a literature (WO 2014/049133 A1)was used instead of 3-cyclopropylpropanal.

¹H-NMR (DMSO-D₆) δ: 4.56-4.58 (1H, m), 4.63-4.65 (2H, m), 4.94-4.96 (2H,m), 7.33 (1H, s), 8.11 (1H, s), 12.54 (1H, br s).

MS (m/z): 209 (M+H)⁺.

Reference Example B-106-[(4-chloropyrimidin-5-yl)oxy]-2,3-difluoro-N,N-di(propan-2-yl)benzamideStep 1 2,3-difluoro-6-methoxy-N,N-di(propan-2-yl)benzamide

To a mixture of 2,3-difluoro-6-methoxybenzoic acid (CAS: 773873-26-0)(2.0 g), diisopropylamine (3.00 mL) and dichloromethane (28 mL) wasadded HATU (4.85 g) under ice-cooling, and the mixture was stirred atroom temperature for 5 hr. Water was added to the reaction solution, andthe mixture was extracted with ethyl acetate. The organic layer waswashed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (2.41 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.14 (6H, d, J=6.7 Hz), 1.55 (6H, d, J=6.7 Hz),3.49-3.56 (1H, m), 3.65-3.72 (1H, m), 3.80 (3H, s), 6.57-6.59 (1H, m),7.05-7.07 (1H, m).

MS (m/z): 272 (M+H)⁺.

Step 2 2,3-difluoro-6-hydroxy-N,N-di(propan-2-yl)benzamide

A mixture of the compound (2.41 g) obtained in the above Step 1 anddichloromethane (17.8 mL) was cooled to −78° C., and boron tribromide(ca. 1 mol/L, dichloromethane solution) (17.8 mL) was added dropwisethereto over 30 min. After the completion of dropwise addition, themixture was stirred at 0° C. for 30 min. After cooled to −78° C.,methanol (5 mL) was added dropwise thereto. After allowed to warm, waterwas added to the reaction solution, and the mixture was extracted withethyl acetate. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure.The residue was purified by silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (2.41 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.37-1.40 (12H, m), 1.62-1.65 (2H, m), 6.61-6.63 (1H,m), 6.97-6.99 (1H, m), 8.01 (1H, s).

MS (m/z): 258 (M+H)⁺.

Step 3 2,3-difluoro-N,N-di(propan-2-yl)-6-[(pyrimidin-5-yl)oxy]benzamide

A mixture of the compound (2.41 g) obtained in the above Step 2,5-bromopyrimidine (CAS: 4595-59-9) (4.24 g), DMF (44.5 mL) and cesiumcarbonate (8.70 g) was stirred at 120° C. for 12 hr. Water was added tothe reaction solution, and the mixture was extracted with ethyl acetate.The organic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate) togive the title compound (812 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.17 (3H, d, J=6.4 Hz), 1.22 (3H, d, J=6.4 Hz), 1.38(3H, d, J=6.7 Hz), 1.52 (3H, d, J=6.7 Hz), 3.48-3.55 (1H, m), 3.74-3.81(1H, m), 6.75-6.78 (1H, m), 7.17-7.19 (1H, m), 8.48 (2H, s), 8.98 (1H,s).

MS (m/z): 336 (M+H)⁺.

Step 42,3-difluoro-6-[(1-oxo-1λ⁵-pyrimidin-5-yl)oxy]-N,N-di(propan-2-yl)benzamide

The compound (810 mg) obtained in the above Step 3 was dissolved indichloromethane (24.2 mL), 3-chloroperbenzoic acid (containing 30%water) (1.79 g) was added thereto under ice-cooling, and the mixture wasstirred at 0° C. for 1 hr, and then at room temperature for 20 hr. Thereaction solution was ice-cooled, and saturated aqueous sodiumthiosulfate solution was added thereto. The organic layer was washedtwice with saturated aqueous sodium hydrogencarbonate solution, driedover anhydrous sodium sulfate, and concentrated under reduced pressureto give the title compound as an oil (849 mg). The obtained compound wasdirectly used in the next reaction.

MS (m/z): 352 (M+H)⁺.

Step 56-[(4-chloropyrimidin-5-yl)oxy]-2,3-difluoro-N,N-di(propan-2-yl)benzamide

To a mixture of TEA (0.502 mL) and chloroform (1.3 mL) was addedphosphorus oxychloride (0.92 g) under ice-cooling. A solution of thecompound (849 mg) obtained in the above Step 4 in chloroform (12 mL) wasadded dropwise thereto. The mixture was heated to 65° C., and stirredfor 4.5 hr. After allowed to cool, the reaction solution was addedlittle by little to ice-cooled saturated aqueous sodiumhydrogencarbonate solution, and the mixture was stirred vigorously for30 min. The mixture was adjusted to pH 7 to 8 with saturated sodiumhydrogencarbonate, and subjected to liquid separation. The aqueous layerwas extracted with dichloromethane, and the organic layers werecombined, dried over anhydrous sodium sulfate, and concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (236mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.22 (3H, d, J=6.4 Hz), 1.25 (3H, d, J=6.4 Hz), 1.39(3H, d, J=7.0 Hz), 1.52 (3H, d, J=7.0 Hz), 3.50-3.57 (1H, m), 3.78-3.80(1H, m), 6.74-6.77 (1H, m), 7.18-7.21 (1H, m), 8.29 (1H, s), 8.75 (1H,s).

MS (m/z): 370, 372 (M+H)⁺.

Reference Example C-1(1R,3S)—N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamineStep 1 benzyl[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate

The compound (11.6 g) obtained in Step 2 of Reference Example A-8 wasdissolved in 2-propanol (407 mL), and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (CAS:1628317-85-0) (10.3 g) synthesized according to the method described ina literature (cancer cell 2015, 27, 589-602.) and DIPEA (21.6 mL) wereadded thereto, and the reaction solution was heated under reflux for 3days. The reaction solution was allowed to cool to room temperature, andconcentrated under reduced pressure. The residue was purified by silicagel chromatography (dichloromethane/ethyl acetate) to give the titlecompound (13.2 g) as a solid.

¹H-NMR (CDCl₃)b: 1.05-1.22 (3H, m), 1.51-1.57 (1H, m), 1.88 (1H, d,J=13.0 Hz), 2.08 (1H, d, J=13.0 Hz), 2.16 (1H, d, J=11.0 Hz), 2.52 (1H,d, J=11.0 Hz), 3.63 (2H, q, J=10.3 Hz), 3.67-3.76 (1H, m), 4.22-4.34(1H, m), 4.59-4.70 (1H, m), 4.91 (1H, d, J=8.0 Hz), 5.10 (2H, s), 7.02(1H, s), 7.38-7.26 (5H, m), 8.47 (1H, s).

MS (m/z): 465 (M+H)⁺.

Step 2(1R,3S)—N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine

The compound (12.9 g) obtained in the above Step 1 was dissolved indichloromethane (92.6 mL). Iodotrimethylsilane (5.33 mL) was addeddropwise thereto under ice-cooling. The reaction solution was warmed toroom temperature, and stirred for 2 hr. Methanol (0.500 mL) was added tothe reaction solution, and the mixture was concentrated under reducedpressure. The residue was purified by amino silica gel chromatography(dichloromethane/methanol) to give the title compound (4.00 g) as anoil.

¹H-NMR (CDCl₃)b: 1.22-1.50 (4H, m), 1.78-1.96 (3H, m), 2.13 (1H, d,J=13.5 Hz), 3.11-3.21 (1H, m), 3.64 (2H, q, J=10.3 Hz), 4.31-4.42 (1H,m), 6.44 (1H, br s), 7.03 (1H, s), 8.46 (1H, s).

MS (m/z): 331 (M+H)⁺.

Reference Example C-2(1R,3S)—N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate

The compound (6.20 g) obtained in Step 2 of Reference Example A-2 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (6.72 g) weredissolved in 2-propanol (90 mL), and DIPEA (9.27 mL) was added thereto,the mixture was heated under reflux for 7 hr. Ethyl acetate andsaturated brine were added to the reaction solution, and the mixture wassubjected to liquid separation, and the aqueous layer was extracted withethyl acetate. The organic layers were combined, washed with saturatedbrine, dried over anhydrous sodium sulfate, and concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (7.50g) as a solid.

¹H-NMR (CDCl₃) δ: 1.04-1.20 (3H, m), 1.44 (9H, s), 1.47-1.56 (1H, m),1.84-1.89 (1H, m), 2.02-2.07 (1H, m), 2.13-2.18 (1H, m), 2.47-2.50 (1H,m), 3.55-3.68 (1H, m), 3.63 (2H, q, J=10.1 Hz), 4.21-4.31 (1H, m), 4.43(1H, br s), 4.97 (1H, d, J=7.3 Hz), 7.03 (1H, s), 8.47 (1H, s).

Step 2(1R,3S)—N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.29-1.44 (3H, m), 1.69-2.02 (4H, m), 2.23-2.29 (1H,m), 3.14 (1H, br s), 4.16 (2H, q, J=10.9 Hz), 4.31 (1H, br s), 8.13 (1H,s), 8.48 (3H, br s), 8.68 (1H, s), 10.01 (1H, d, J=7.9 Hz).

Reference Example C-3(1R,3R)—N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1R,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using tert-butyl[(1R,3R)-3-aminocyclopentyl]carbamate (CAS: 1009075-44-8) and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (CDCl₃) δ: 1.45 (9H, s), 1.46-1.63 (2H, m), 1.91-2.05 (2H, m),2.18-2.25 (1H, m), 2.30-2.39 (1H, m), 3.62 (2H, q, J=10.4 Hz), 4.13-4.20(1H, m), 4.63-4.71 (2H, m), 5.54 (1H, d, J=4.9 Hz), 7.11 (1H, s), 8.45(1H, s).

MS (m/z): 417 (M+H)⁺.

Step 2(1R,3R)—N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.61-1.79 (2H, m), 2.11 (2H, t, J=7.3 Hz), 2.14-2.27(2H, m), 3.70-3.78 (1H, m), 4.14 (2H, q, J=10.9 Hz), 4.72-4.80 (1H, m),7.95 (1H, s), 8.19 (3H, br s), 8.58 (1H, s), 9.11 (1H, br s).

MS (m/z): 317 (M+H)⁺.

Reference Example C-4(1R,3S)—N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in

Reference Example A-4 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine

¹H-NMR (CDCl₃) δ: 1.46 (9H, s), 1.57-1.68 (2H, m), 1.87-2.11 (3H, m),2.36-2.43 (1H, m), 3.27 (3H, s), 3.64 (2H, q, J=10.5 Hz), 4.00 (1H, brs), 4.95 (1H, br s), 5.10 (1H, br s), 7.32 (1H, s), 8.43 (1H, s).

MS (m/z):431 (M+H)⁺.

Step 2(1R,3S)—N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.82-2.14 (5H, m), 2.28-2.35 (1H, m), 3.38 (3H, s),3.53-3.62 (1H, m), 4.17 (2H, q, J=10.9 Hz), 5.24-5.35 (1H, m), 7.88 (1H,s), 8.50 (3H, br s), 8.64 (1H, s).

Reference Example C-5(1R,3S)—N¹-ethyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{ethyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 2 ofReference Example A-5 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (CDCl₃) δ: 1.38 (3H, t, J=7.3 Hz), 1.47 (9H, s), 1.71-1.82 (2H,m), 1.88-1.97 (2H, m), 2.06-2.19 (1H, m), 2.33-2.40 (1H, m), 3.64 (2H,q, J=10.3 Hz), 3.65-3.75 (2H, m), 4.07 (1H, br s), 4.55 (1H, br s), 5.83(1H, s), 7.20 (1H, s), 8.43 (1H, s).

MS (m/z): 445 (M+H)⁺.

Step 2(1R,3S)—N¹-ethyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.28 (3H, t, J=6.7 Hz), 1.79-2.11 (5H, m), 2.29-2.37(1H, m), 3.57-3.66 (1H, m), 3.71-3.85 (2H, m), 4.16 (2H, q, J=10.7 Hz),5.05-5.14 (1H, m), 7.63 (1H, s), 8.26 (3H, br s), 8.49 (1H, s).

Reference Example C-6(1R,3S)—N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]amino}cyclopentyl]carbamate

A mixture of the compound (0.0974 g) obtained in Step 7 of ReferenceExample B-7, BOP (CAS: 56602-33-6) (0.250 g), acetonitrile (8.5 mL) andDBU (CAS: 6674-22-2) (0.145 mL) was stirred at room temperature for 7min. Then, a solution of the compound (0.218 g) obtained in ReferenceExample A-4 in acetonitrile (8.5 mL) was added thereto, and the mixturewas stirred at 60° C. for 1 hr. The reaction solution was concentratedunder reduced pressure, and the obtained residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (0.178 g) as an oil.

¹H-NMR (DMSO-D₆) δ: 1.39 (9H, s), 1.53-1.63 (1H, m), 1.65-1.76 (1H, m),1.80-1.95 (3H, m), 2.24-2.34 (1H, m), 3.19 (3H, s), 3.76-3.87 (1H, m),3.89 (2H, q, J=11.5 Hz), 4.75-4.86 (1H, m), 7.05 (1H, d, J=7.3 Hz), 7.75(2H, s), 8.05 (1H, br s), 8.53 (1H, s).

MS (m/z): 425 (M+H)⁺.

Step 2(1R,3S)—N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.83-2.22 (5H, m), 2.38-2.49 (1H, m), 3.51 (3H, s),3.97 (2H, q, J=11.3 Hz), 5.21-5.35 (1H, m), 7.92 (1H, d, J=8.5 Hz), 7.99(1H, d, J=8.5 Hz), 8.31 (1H, br s), 8.35 (3H, br s), 8.80-8.84 (1H, m).

MS (m/z): 325 (M+H)⁺.

Reference Example C-7 N⁴-[(1R,3S)-3-aminocyclopentyl]-N²,N⁴-dimethyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine-2,4-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Reference ExampleA-4 and the compound obtained in Step 2 of Reference Example B-4.

¹H-NMR (CDCl₃) δ: 7.29 (1H, s), 5.09 (1H, br s), 4.72 (1H, br s),4.06-3.93 (1H, m), 3.64 (1H, d, J=10.5 Hz), 3.59 (1H, d, J=10.5 Hz),3.28 (3H, s), 2.46-2.37 (1H, m), 2.13-1.95 (2H, m), 1.94-1.81 (1H, m),1.71-1.59 (2H, m), 1.46 (9H, s).

MS (m/z): 465, 467 (M+H)⁺.

Step 2 tert-butyl[(1S,3R)-3-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The compound (800 mg) obtained in the above Step 1 was suspended inbutyronitrile (6 mL), and 40% aqueous methylamine solution (0.741 mL)was added thereto, and the mixture was stirred with heating in amicrowave reactor at 150° C. for 1 hr and 45 min. Ethyl acetate andsaturated brine were added to the reaction solution, and the mixture wassubjected to liquid separation. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. A mixed solvent of dichloromethane/n-hexane=1/3was added to the obtained residue, and the solid was collected byfiltration to give the title compound (650 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.39 (9H, s), 1.52-1.61 (2H, m), 1.71-1.88 (3H, m),2.09-2.16 (1H, m), 2.76 (3H, d, J=4.9 Hz), 3.10 (3H, s), 3.76-3.85 (1H,m), 3.88 (2H, q, J=11.0 Hz), 5.00-5.09 (1H, m), 6.60 (1H, q, J=4.9 Hz),7.03 (1H, d, J=7.9 Hz), 7.38 (1H, s).

MS (m/z): 460 (M+H)⁺.

Step 3N⁴-[(1R,3S)-3-aminocyclopentyl]-N²,N⁴-dimethyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine-2,4-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (DMSO-D₆) δ: 1.84-1.94 (3H, m), 2.01-2.08 (2H, m), 2.31-2.37 (1H,m), 2.94 (3H, s), 3.34 (3H, s), 3.52-3.60 (1H, m), 4.05 (2H, q, J=11.0Hz), 5.18 (1H, br s), 7.66 (1H, s), 8.02 (1H, br s), 8.47 (3H, s).

MS (m/z): 360 (M+H)⁺.

Reference Example C-8(1R,3S)—N¹-[6-(methoxymethyl)thieno[2,3-d]pyrimidin-4-yl]-N¹-methylcyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{[6-(methoxymethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-6, using the compound obtained in Reference ExampleA-4 and the compound obtained in Reference Example B-8.

¹H-NMR (CDCl₃) δ: 1.28-1.29 (1H, m), 1.49 (9H, s), 1.93-2.14 (4H, m),2.41-2.43 (1H, m), 3.29 (3H, s), 3.46 (3H, s), 4.01-4.04 (1H, m), 4.68(2H, s), 5.09-5.11 (1H, m), 7.29 (1H, s), 7.31 (1H, s), 8.44 (1H, s).

MS (m/z): 393 (M+H)⁺.

Step 2(1R,3S)—N¹-[6-(methoxymethyl)thieno[2,3-d]pyrimidin-4-yl]-N¹-methylcyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound (140 mg) obtained in the aboveStep 1.

MS (m/z): 293 (M+H)⁺.

Reference Example C-92-{[(1R,3S)-3-aminocyclopentyl][6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}ethan-1-olhydrochloride Step 1 tert-butyl[(1S,3R)-3-{(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 2 ofReference Example A-19 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (DMSO-D₆) δ: −0.03 (6H, s), 0.81 (9H, s), 1.17-1.22 (1H, m), 1.39(9H, s), 1.57-1.59 (1H, m), 1.66-1.72 (1H, m), 1.85-1.88 (2H, m),2.20-2.22 (1H, m), 3.71-3.86 (5H, m), 4.02-4.13 (2H, m), 4.81-4.83 (1H,m), 7.08 (1H, d, J=7.4 Hz), 7.55 (1H, s), 8.36 (1H, s).

MS (m/z): 575 (M+H)⁺.

Step 22-{[(1R,3S)-3-aminocyclopentyl][6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}ethan-1-olhydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.The obtained compound was directly used in the next step withoutpurification.

MS (m/z): 361 (M+H)⁺.

Reference Example C-10(1R,3S)—N¹-methyl-N¹-[2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{methyl[2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Reference ExampleA-4 and the compound obtained in Step 3 of Reference Example B-1.

¹H-NMR (CDCl₃) δ: 1.48 (9H, s), 1.59-1.62 (2H, m), 1.92-2.02 (3H, m),2.36-2.43 (1H, m), 2.57 (3H, s), 3.24 (3H, s), 3.60 (2H, q, J=10.1 Hz),4.00 (1H, br s), 4.74 (1H, br s), 5.15-5.17 (1H, br m), 7.26 (1H, s).

MS (m/z): 445 (M+H)⁺.

Step 2(1R,3S)—N¹-methyl-N¹-[2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.76-1.96 (3H, m), 1.96-2.11 (2H, m), 2.24-2.35 (1H,m), 2.52 (3H, s), 3.33 (3H, s), 3.58-3.64 (1H, m), 4.11 (2H, q, J=11.1Hz), 5.31-5.33 (1H, br m), 7.76 (1H, s), 8.23 (3H, br s).

MS (m/z): 345 (M+H)⁺.

Reference Example C-11(1R,3S)—N¹-methyl-N¹-[6-(oxetan-3-yl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,3R)-3-{methyl[6-(oxetan-3-yl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-6, using the compound obtained in Step 1 ofReference Example A-4 and the compound obtained in Reference ExampleB-9.

¹H-NMR (DMSO-D₆) δ: 1.39 (9H, s), 1.56-1.64 (2H, m), 1.78-1.90 (3H, m),2.11-2.17 (1H, m), 3.20 (3H, s), 3.82-3.84 (1H, m), 4.60-4.69 (3H, m),4.96-4.98 (2H, m), 5.14-5.16 (1H, m), 7.04 (1H, d, J=7.4 Hz), 7.50 (1H,s), 8.32 (1H, s).

MS (m/z): 405 (M+H)⁺.

Step 2(1R,3S)—N¹-methyl-N¹-[6-(oxetan-3-yl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.The obtained compound was directly used in the next step withoutpurification.

¹H-NMR (DMSO-D₆) δ: 1.80-2.33 (7H, m), 3.35 (3H, s), 3.67-3.71 (2H, m),3.78-3.85 (1H, m), 3.88-3.95 (1H, m), 4.01-4.04 (1H, m), 5.27-5.30 (1H,m), 7.65 (1H, s), 8.26 (3H, s), 8.54 (1H, s).

MS (m/z): 305 (M+H)⁺.

Reference Example C-12(1R,3S)—N¹-(6-cyclopropylthieno[2,3-d]pyrimidin-4-yl)-N¹-methylcyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl{(1S,3R)-3-[(6-cyclopropylthieno[2,3-d]pyrimidin-4-yl)(methyl)amino]cyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Reference ExampleA-4 and the compound obtained in Reference Example B-3.

¹H-NMR (DMSO-D₆) δ: 0.79-0.80 (2H, m), 1.04-1.07 (2H, m), 1.40 (9H, s),1.52-1.68 (2H, m), 1.78-1.94 (3H, m), 2.16-2.23 (2H, m), 3.18 (3H, s),3.78-3.86 (1H, m), 5.04-5.12 (1H, m), 6.68 (1H, s), 7.24 (1H, s), 8.25(1H, s).

MS (m/z): 389 (M+H)⁺.

Step 2(1R,3S)—N¹-(6-cyclopropylthieno[2,3-d]pyrimidin-4-yl)-N¹-methylcyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.The obtained compound was directly used in the next step withoutpurification.

MS (m/z): 289 (M+H)⁺.

Reference Example C-13(1S,3R)-5,5-difluoro-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride Step 1 tert-butyl[(1R,5S)-3,3-difluoro-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 2 ofReference Example A-18 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (CDCl₃) δ: 1.43-1.45 (11H, m), 1.66-1.78 (2H, m), 2.51-2.58 (2H,m), 3.59-3.62 (2H, m), 3.85-3.87 (1H, m), 4.66-4.68 (2H, m), 5.66-5.68(1H, m), 7.11 (1H, s), 8.48 (1H, s).

MS (m/z): 467 (M+H)⁺.

Step 2(1S,3R)-5,5-difluoro-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (CD₃OD) δ: 1.84-2.23 (3H, m), 2.50-2.53 (3H, m), 3.52-3.58 (1H,m), 4.04 (2H, q, J=10.4 Hz), 4.72-4.75 (1H, m), 7.86 (1H, s), 8.76 (1H,s).

MS (m/z): 367 (M+H)⁺.

Reference Example C-14(1R,2S,4R)-4-amino-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride Step 1 benzyl[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

A mixture of the compound (43.3 g) obtained in Step 14 of ReferenceExample A-1, 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(CAS: 1628317-85-0) (43.5 g), DIPEA (57.1 mL) and 2-propanol (820 mL)was stirred at 90° C. for 5 hr. The reaction solution was concentratedunder reduced pressure, water was added thereto, and the mixture wasextracted with ethyl acetate/dichloromethane/methanol. The organic layerwas dried over anhydrous sodium sulfate, and concentrated under reducedpressure. Ethyl acetate/n-hexane was added to the residue, and the solidwas collected by filtration to give the title compound (64.2 g) as asolid. The filtrate was concentrated, and ethyl acetate/n-hexane wasadded to the residue, and the solid was collected by filtration to givethe title compound (26.5 g) as a solid. The filtrate was concentrated,and the residue was purified by silica gel column chromatography(dichloromethane/ethyl acetate). Ethyl acetate was added to the obtainedsolid, and the solid was collected by filtration to give the titlecompound (3.67 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.72-1.84 (1H, m), 2.22-2.44 (3H, m), 3.49 (3H, s),3.62 (2H, q, J=10.2 Hz), 3.90-3.98 (1H, m), 4.08-4.19 (1H, m), 4.54-4.62(1H, m), 4.65-4.73 (1H, m), 5.12 (2H, s), 5.46 (1H, d, J=7.9 Hz),7.31-7.42 (6H, m), 8.35 (1H, s).

MS (m/z): 481 (M+H)⁺.

Step 2(1R,2S,4R)-4-amino-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

To a mixture of the compound (8.98 g) obtained in the above Step 1 andacetonitrile (90 mL) was added iodotrimethylsilane (8.1 mL) at 0° C.,and the mixture was stirred at 0° C. for 30 min. 1N Hydrochloric acidand water were added to the reaction solution, and the mixture waswashed with ethyl acetate. The organic layer was extracted with 1Nhydrochloric acid, 2N aqueous sodium hydroxide solution was added to thecombined aqueous layers, and the mixture was extracted nine times withdichloromethane/methanol. The combined organic layers were dried overanhydrous sodium sulfate, and concentrated under reduced pressure togive the title compound (6.53 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.65-1.73 (1H, m), 1.80-1.91 (1H, m), 2.03-2.12 (1H,m), 2.25-2.36 (1H, m), 3.56 (3H, s), 3.58-3.73 (3H, m), 4.46-4.52 (1H,m), 5.06-5.16 (1H, m), 7.41 (1H, s), 8.39 (1H, s).

MS (m/z): 347 (M+H)⁺.

Step 3(1R,2S,4R)-4-amino-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

A mixture of the compound (6.53 g) obtained in the above Step 2, 1Nhydrochloric acid (21 mL) and acetonitrile (60 mL) was stirred at roomtemperature for 10 min. The reaction solution was concentrated underreduced pressure, acetonitrile was added thereto, and the solid wascollected by filtration to give the title compound (5.83 g) as a solid.The filtrate was concentrated, acetonitrile was added thereto, and thesolid was collected by filtration to give the title compound (1.13 g) asa solid.

¹H-NMR (DMSO-D₆) δ: 1.59-1.69 (1H, m), 2.18-2.27 (2H, m), 2.32-2.43 (1H,m), 3.43 (3H, s), 3.50-3.63 (1H, m), 4.10 (2H, q, J=11.1 Hz), 4.31-4.39(1H, m), 4.89-5.00 (1H, m), 5.19 (1H, br s), 7.75 (1H, s), 8.08 (3H, brs), 8.38 (1H, s).

MS (m/z): 347 (M+H)⁺.

Reference Example C-15(1R,2S,4R)-4-amino-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride Step 1 tert-butyl[(1R,3R,4S)-3-hydroxy-4-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]carbamate

To the compound (3.33 g) obtained in Step 2 of Reference Example A-15and the compound (4.09 g) obtained in Step 4 of Reference Example B-5were added 2-propanol (49 mL) and DIPEA (7.56 mL), and the mixture washeated under reflux for 3 hr, and allowed to stand overnight at roomtemperature. Ethyl acetate and saturated brine were added to thereaction solution, and the mixture was subjected to liquid separation.The organic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (5.05 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.46 (9H, s), 1.73 (1H, d, J=13.4 Hz), 2.23-2.30 (2H,m), 2.35-2.42 (1H, m), 3.48 (3H, s), 3.56 (2H, q, J=10.1 Hz), 3.67 (1H,br s), 3.93 (3H, s), 3.95-4.03 (1H, m), 4.54-4.58 (1H, m), 4.72 (1H, td,J=9.7, 4.9 Hz), 5.13-5.18 (1H, m), 7.27 (1H, s).

MS (m/z): 477 (M+H)⁺.

Step 2(1R,2S,4R)-4-amino-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

A mixture of the compound (810 mg) obtained in the above Step 1,dichloromethane (7 mL) and hydrogen chloride (4 mol/L, 1,4-dioxanesolution, 7 mL) was stirred at room temperature. A small amount ofmethanol was added thereto to make a solution, and the solution wasfurther stirred, and concentrated under reduced pressure. The residuewas suspended in n-hexane/ethanol, and the solid was collected byfiltration to give the title compound (724 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.64-1.68 (1H, m), 2.23-2.26 (2H, m), 2.34-2.42 (1H,m), 3.40 (3H, s), 3.54-3.58 (1H, m), 3.65-3.68 (1H, m), 3.86 (3H, s),3.97-4.09 (2H, m), 4.34-4.36 (1H, m), 4.83-4.85 (1H, m), 7.64 (1H, s),8.20-8.23 (3H, m).

MS (m/z): 377 (M+H)⁺.

Reference Example C-16(1S,2S,4R)-4-amino-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olStep 1 benzyl[(1R,3S,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 9 ofReference Example A-14 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (CDCl₃) δ: 1.85-1.99 (2H, m), 2.16-2.23 (1H, m), 2.47 (1H, br s),3.32 (3H, s), 3.66 (2H, q, J=10.2 Hz), 4.23-4.35 (2H, m), 4.65 (1H, dt,J=10.4, 7.4 Hz), 4.95 (1H, br s), 5.12 (3H, br s), 7.32-7.42 (5H, m),8.43 (1H, s), 7.37 (1H, s).

MS (m/z): 481 (M+H)⁺.

Step 2(1S,2S,4R)-4-amino-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.28-1.39 (1H, m), 1.59-1.73 (4H, m), 2.15-2.24 (1H,m), 3.24 (3H, s), 3.29-3.42 (2H, m), 4.06 (1H, d, J=11.0 Hz), 4.12 (1H,d, J=11.0 Hz), 4.33-4.42 (1H, m), 4.80-4.93 (2H, m), 7.73 (1H, s), 8.33(1H, s).

MS (m/z): 347 (M+H)⁺.

Reference Example C-17(1R,3S,5R)-3-amino-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamide(racemate) Step 1 benzyl[(1S,3S,5R)-3-(dimethylcarbamoyl)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate(racemate)

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 6 ofReference Example A-7 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (CDCl₃) δ: 1.37 (1H, q, J=11.5 Hz), 1.57-1.71 (2H, m), 1.91 (1H,s), 2.09-2.13 (2H, m), 2.42-2.45 (1H, m), 2.89-2.91 (1H, m), 2.94 (3H,s), 3.12 (3H, s), 3.60 (2H, q, J=10.3 Hz), 3.75-3.78 (1H, m), 4.45-4.48(1H, m), 5.06 (2H, s), 5.28-5.30 (1H, m), 6.34 (1H, d, J=7.9 Hz),7.29-7.32 (5H, m), 8.46 (1H, s).

MS (m/z): 536 (M+H)⁺.

Step 2(1R,3S,5R)-3-amino-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamide(racemate)

The title compound was obtained in the same manner as in Step 2 ofReference Example C-1, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.21 (1H, q, J=11.5 Hz), 1.42 (1H, q, J=12.4 Hz), 1.62(1H, q, J=12.1 Hz), 1.95 (1H, d, J=13.4 Hz), 2.16 (1H, d, J=12.8 Hz),2.31 (1H, d, J=11.5 Hz), 2.86-2.89 (1H, m), 2.98 (3H, s), 3.00-3.02 (1H,m), 3.13 (3H, s), 3.64 (2H, q, J=10 Hz), 4.41-4.44 (1H, m), 5.96 (1H, d,J=8.5 Hz), 7.24 (1H, s), 8.47 (1H, s).

MS (m/z): 402 (M+H)⁺.

Reference Example C-18(1S,3R,5R)-5-methoxy-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine(racemate) Step 1 benzyl[(1R,3S,5S)-3-methoxy-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate(racemate)

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 4 ofReference Example A-11 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

MS (m/z): 495 (M+H)⁺.

Step 2(1S,3R,5R)-5-methoxy-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine(racemate)

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 1.

¹H-NMR (CD₃OD) δ: 1.06 (1H, q, J=11.6 Hz), 1.17-1.29 (2H, m), 2.20 (1H,d, J=11.6 Hz), 2.30 (1H, d, J=12.2 Hz), 2.42 (1H, d, J=11.0 Hz),2.80-2.88 (1H, m), 3.37-3.44 (4H, m), 3.85 (2H, q, J=10.4 Hz), 4.21-4.29(1H, m), 7.51 (1H, s), 8.31 (1H, s).

MS (m/z):361 (M+H)⁺.

Reference Example C-19(1R,3R,5S)-3-amino-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-ol(racemate) Step 1 benzyl[(1R,3S,5S)-3-(methoxymethoxy)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate(racemate)

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 5 ofReference Example A-6 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine.

¹H-NMR (CDCl₃) δ: 1.31-1.47 (3H, m), 2.34-2.43 (3H, m), 3.38 (3H, s),3.60-3.62 (2H, m), 3.80-3.85 (2H, m), 4.40-4.41 (1H, m), 4.72-4.79 (3H,m), 5.10-5.18 (3H, m), 7.01 (1H, s), 7.31-7.36 (5H, m), 8.48 (1H, s).

MS (m/z): 525 (M+H)⁺.

Step 2(1R,3R,5S)-3-amino-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-ol(racemate)

The title compound was obtained in the same manner as in Step 2 ofReference Example C-1, using the compound obtained in the above Step 1.

¹H-NMR (CD₃OD) δ: 1.15-1.19 (2H, m), 1.33-1.36 (1H, m), 2.16-2.19 (2H,m), 2.25-2.28 (1H, m), 2.83-2.86 (1H, m), 3.72-3.75 (1H, m), 3.84-3.87(2H, m), 4.25-4.27 (1H, m), 7.52 (1H, s), 8.30 (1H, s).

MS (m/z): 347 (M+H)⁺.

Reference Example C-20(1R,2S,4R)-4-amino-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olStep 1 benzyl[(1R,3S,4R)-3-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-hydroxycyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 14 ofReference Example A-1 and the compound obtained in Step 2 of ReferenceExample B-4.

¹H-NMR (CDCl₃) δ: 1.76-1.80 (1H, m), 2.29-2.33 (2H, m), 2.39-2.46 (1H,m), 3.14 (1H, d, J=5.5 Hz), 3.50 (3H, s), 3.61 (2H, q, J=10.2 Hz),4.06-4.11 (1H, m), 4.55-4.58 (1H, m), 4.80 (1H, td, J=10.0, 4.7 Hz),5.13 (2H, s), 5.39 (1H, d, J=6.7 Hz), 7.33-7.38 (6H, m).

Step 2 benzyl[(1R,3R,4S)-3-hydroxy-4-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 2 ofReference Example C-7, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.71-1.76 (1H, m), 2.24-2.32 (3H, m), 2.92 (3H, d,J=5.5 Hz), 3.40 (3H, s), 3.51 (2H, q, J=10.4 Hz), 4.12 (2H, q, J=7.2Hz), 4.43-4.47 (1H, m), 4.54-4.56 (1H, m), 4.74-4.77 (1H, m), 5.11 (2H,s), 5.38-5.42 (1H, m), 7.13 (1H, s), 7.30-7.41 (5H, m).

MS (m/z): 510 (M+H)⁺.

Step 3(1R,2S,4R)-4-amino-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 2.

1H-NMR (CDCl₃) δ: 1.62-1.66 (1H, m), 1.85-1.92 (1H, m), 2.07-2.12 (1H,m), 2.21-2.28 (1H, m), 2.96 (3H, d, J=5.5 Hz), 3.46 (3H, s), 3.52 (2H,q, J=10.4 Hz), 3.58-3.62 (1H, m), 4.47-4.50 (1H, m), 4.71-4.74 (1H, m),4.84 (1H, td, J=9.7, 5.1 Hz), 7.19 (1H, s).

Reference Example C-21(1R,2S,4R)-4-amino-2-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride Step 1 tert-butyl[(1R,3S,4R)-3-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-hydroxycyclopentyl]carbamate

A mixture of the compound (8.41 g) obtained in Step 1 of ReferenceExample C-20, acetonitrile (100 mL) and iodotrimethylsilane (6.71 mL)was stirred under ice-cooling for 1 hr. Methanol (10 mL) was addedthereto, the mixture was stirred for 20 min, and the solvent wasevaporated under reduced pressure. THF (100 mL), water (100 mL), sodiumcarbonate (6.93 g) and di-tert-butyl dicarbonate (5.70 g) were added tothe obtained oil, and the mixture was stirred at room temperature for 1hr. Water and ethyl acetate were added to the reaction solution, and themixture was subjected to liquid separation. The organic layer was washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. Dichloromethane/n-hexane was addedto the obtained residue, and the solid was collected by filtration togive the title compound (6.51 g) as a solid. The filtrate wasconcentrated, and the residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound(0.903 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.47 (9H, s), 1.74-1.77 (1H, m), 2.17-2.31 (2H, m),2.39-2.46 (1H, m), 3.52 (3H, s), 3.57-3.69 (3H, m), 3.94-3.99 (1H, m),4.53-4.55 (1H, m), 4.76-4.79 (1H, m), 5.13-5.15 (1H, m), 7.36 (1H, s).

MS (m/z): 481 (M+H)+.

Step 2(1R,2S,4R)-4-amino-2-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.67-1.70 (1H, m), 2.23-2.44 (3H, m), 3.41 (3H, s),3.54-3.60 (1H, m), 4.08-4.16 (2H, m), 4.33-4.34 (1H, m), 4.79-4.81 (1H,m), 7.77 (1H, s), 8.29 (3H, s).

MS (m/z): 381 (M+H)+.

Reference Example C-22(1S,2S,4R)-4-amino-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olStep 1 benzyl[(1R,3S,4S)-3-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-hydroxycyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 9 ofReference Example A-14 and the compound obtained in Step 2 of ReferenceExample B-4.

¹H-NMR (CDCl₃) δ: 1.90-2.03 (2H, m), 2.14-2.21 (1H, m), 2.44-2.52 (1H,m), 3.35 (3H, s), 3.63 (2H, q, J=10.0 Hz), 3.96-4.05 (1H, m), 4.21-4.32(1H, m), 4.34-4.43 (1H, m), 4.54-4.61 (1H, m), 4.92-4.99 (1H, m), 5.12(2H, s), 7.31-7.40 (6H, m).

Step 2 benzyl[(1R,3S,4S)-3-hydroxy-4-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 2 ofReference Example C-7, using the compound obtained in the above Step 1.

1H-NMR (DMSO-D₆) δ: 1.58-1.66 (1H, m), 1.80-1.85 (2H, m), 2.23-2.31 (1H,m), 2.80 (3H, d, J=4.9 Hz), 3.07 (3H, s), 3.82 (2H, q, J=11.2 Hz),3.98-4.06 (1H, m), 4.25-4.33 (1H, m), 4.62-4.69 (1H, m), 4.73-4.76 (1H,m), 5.03 (2H, s), 6.27-6.33 (1H, m), 7.11-7.20 (1H, m), 7.27-7.35 (5H,m), 7.38 (1H, s).

Step 3(1S,2S,4R)-4-amino-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 2.

1H-NMR (DMSO-D₆) δ: 1.26-1.34 (1H, m), 1.60-1.70 (2H, m), 2.13-2.23 (1H,m), 2.77 (3H, d, J=4.9 Hz), 3.14 (3H, s), 3.30-3.37 (1H, m), 3.89 (2H,q, J=11.4 Hz), 4.30-4.38 (1H, m), 4.67-4.75 (1H, m), 4.84 (1H, d, J=4.9Hz), 6.56-6.59 (1H, m), 7.41 (1H, s).

Reference Example C-23(1R,2S,4R)-4-amino-2-{[2-{[(4-methoxyphenyl)methyl]amino}-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olStep 1 benzyl[(1R,3R,4S)-3-hydroxy-4-{[2-{[(4-methoxyphenyl)methyl]amino}-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]carbamate

A mixture of the compound (1.57 g) obtained in Step 1 of ReferenceExample C-20, 4-methoxybenzylamine (1.97 mL), DIPEA (2.66 mL) andbutyronitrile (9.0 mL) was stirred with heating in a microwave reactorat 150° C. for 1.5 hr. The insoluble substance in the reaction solutionwas filtered off, water was added to the filtrate, and the mixture wasextracted with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure, and the residue was subjected to silica gelcolumn chromatography (chloroform/methanol) to give the title compound(1.62 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.55-1.68 (2H, m), 2.12-2.29 (2H, m), 3.37 (3H, d,J=3.7 Hz), 3.51 (2H, qd, J=10.4, 3.7 Hz), 3.77 (3H, d, J=1.2 Hz),4.00-4.10 (1H, m), 4.38 (1H, br s), 4.42-4.56 (3H, m), 5.09 (2H, br s),5.11-5.20 (1H, m), 5.31 (1H, br s), 6.83 (2H, dd, J=8.6, 2.5 Hz), 7.14(1H, d, J=6.1 Hz), 7.19-7.23 (2H, m), 7.28-7.38 (5H, m).

MS (m/z): 616 (M+H)⁺.

Step 2(1R,2S,4R)-4-amino-2-{[2-{[(4-methoxyphenyl)methyl]amino}-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.54-1.60 (1H, m), 1.80 (1H, ddd, J=14.1, 9.2, 4.9Hz), 2.00 (1H, dt, J=14.1, 5.5 Hz), 2.17 (1H, ddd, J=13.5, 9.2, 6.1 Hz),3.44 (3H, s), 3.51 (2H, q, J=10.0 Hz), 3.53-3.57 (1H, m), 3.79 (3H, s),4.32 (1H, s), 4.52 (2H, d, J=5.5 Hz), 4.81 (1H, td, J=9.7, 5.1 Hz), 5.09(1H, br s), 6.84 (2H, d, J=8.6 Hz), 7.19 (1H, s), 7.24 (2H, d, J=8.6Hz).

MS (m/z): 482 (M+H)⁺.

Reference Example C-242-[(4-{[(1S,2R,4R)-4-amino-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-5-fluoro-N,N-di(propan-2-yl)benzamideStep 1 benzyl{(1R,3S,4R)-3-[(5-{2-[di(propan-2-yl)carbamoyl]-4-fluorophenoxy}pyrimidin-4-yl)(methyl)amino]-4-hydroxycyclopentyl}carbamate

A mixture of the compound (0.233 g) obtained in Step 14 of ReferenceExample A-1,2-[(4-chloropyrimidin-5-yl)oxy]-5-fluoro-N,N-di(propan-2-yl)benzamide(0.308 g) produced according to the method described in a literature (WO2017/214367), 2-propanol (3.50 mL) and DIPEA (0.305 mL) was stirred in amicrowave reactor at 110° C. for 30 min, and then stirred under the samecondition for 2 hr. The compound (0.0463 g) obtained in Step 14 ofReference Example A-1 was added thereto, and the mixture was stirred at110° C. for 2 hr in a microwave reactor, and allowed to stand at roomtemperature for 16 hr. The compound (0.0463 g) obtained in Step 14 ofReference Example A-1 was added thereto, and the mixture was stirred at110° C. for 2 hr in a microwave reactor. The reaction solution wasconcentrated under reduced pressure, diluted with water, and extractedwith ethyl acetate. The organic layer was washed with saturated brine,and dried over anhydrous sodium sulfate. The solvent was evaporatedunder reduced pressure, and the obtained residue was subjected to silicagel column chromatography (n-hexane/ethyl acetate, followed by ethylacetate/methanol) to give the title compound (0.475 g) as a solid.

¹H-NMR (CDCl₃) δ: 0.97-1.77 (14H, m), 2.06-2.39 (3H, m), 3.16-3.26 (3H,m), 3.37-3.54 (1H, m), 3.68-3.88 (1H, m), 3.96-4.12 (1H, m), 4.17-4.55(2H, m), 5.09 (2H, s), 5.29-5.43 (1H, m), 6.62-7.07 (3H, m), 7.28-7.41(5H, m), 7.79-7.91 (1H, m), 8.37-8.44 (1H, m).

MS (m/z): 580 (M+H)⁺.

Step 22-[(4-{[(1S,2R,4R)-4-amino-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-5-fluoro-N,N-di(propan-2-yl)benzamide

The title compound was obtained in the same manner as in Step 2 ofReference Example A-2, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.01-1.22 (7H, m), 1.31-1.59 (8H, m), 1.89-1.99 (1H,m), 2.11-2.34 (3H, m), 3.17-3.23 (3H, m), 3.40-3.55 (1H, m), 3.66-3.87(2H, m), 4.31-4.37 (1H, m), 4.55-4.71 (1H, m), 6.51-6.70 (1H, m),6.85-7.01 (2H, m), 7.85-7.88 (1H, m), 8.44 (1H, s).

MS (m/z): 446 (M+H)⁺.

Reference Example C-25(1R,2S,4R)-4-amino-2-[(5-{4-fluoro-2-[4-(propan-2-yl)pyrimidin-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-olStep 1 benzyl[(1R,3S,4R)-3-{[5-(2-bromo-4-fluorophenoxy)pyrimidin-4-yl](methyl)amino}-4-hydroxycyclopentyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 14 ofReference Example A-1 and 5-(2-bromo-4-fluorophenoxy)-4-chloropyrimidineproduced according to the method described in a literature (WO2017/214367).

¹H-NMR (CDCl₃) δ: 1.65-1.69 (1H, m), 2.19-2.27 (3H, m), 3.14-3.20 (1H,m), 3.25 (3H, s), 4.08-4.12 (1H, m), 4.39-4.41 (2H, m), 5.10 (2H, s),5.23-5.26 (1H, m), 6.68-6.72 (1H, m), 6.96-6.99 (1H, m), 7.34-7.40 (5H,m), 7.83 (1H, s), 8.42 (1H, s).

MS (m/z): 531, 533 (M+H)⁺.

Step 2 benzyl{(1R,3S,4R)-3-[(5-{4-fluoro-2-[4-(propan-2-yl)pyrimidin-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]-4-hydroxycyclopentyl}carbamate

A mixture of the compound (140 mg) obtained in the above Step 1,4-isopropylpyrimidine-5-boronic acid (CAS: 913835-27-5) (52.5 mg),tetrakis(triphenylphosphine)palladium(0) (30.4 mg), sodium carbonate(112 mg), water (1.1 mL) and 1,4-dioxane (3.3 mL) was stirred at 90° C.for 1.5 hr. Water was added to the reaction solution, and the mixturewas extracted with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was subjected to silica gel columnchromatography (n-hexane/ethyl acetate). n-Hexane/ethyl acetate wasadded to the obtained residue, and the resulting solid was collected byfiltration to give the title compound (92.5 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.22-1.26 (6H, m), 1.66-1.69 (1H, m), 2.12-2.23 (3H,m), 3.01-3.07 (4H, m), 3.49-3.50 (1H, m), 3.99-4.06 (1H, m), 4.34-4.37(1H, m), 5.10 (2H, s), 5.30 (1H, s), 6.81 (1H, s), 7.01-7.03 (1H, m),7.09-7.14 (1H, m), 7.30-7.39 (5H, m), 7.79-7.82 (1H, m), 8.33-8.35 (1H,m), 8.47 (1H, s), 9.15 (1H, s).

MS (m/z): 573 (M+H)⁺.

Step 3(1R,2S,4R)-4-amino-2-[(5-{4-fluoro-2-[4-(propan-2-yl)pyrimidin-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 2.

¹H-NMR (DMSO-D₆) δ: 1.15-1.21 (7H, m), 1.71-1.76 (3H, m), 1.98-2.01 (1H,m), 2.32-2.34 (1H, m), 2.99-3.01 (5H, m), 4.01-4.08 (1H, m), 4.33-4.35(1H, m), 6.89-6.91 (1H, m), 7.30-7.32 (1H, m), 7.39-7.41 (1H, m),7.90-7.94 (1H, m), 8.33 (1H, s), 8.64-8.67 (1H, m), 9.17 (1H, s).

MS (m/z): 439 (M+H)⁺.

Reference Example C-26(1R,2S,4R)-4-amino-2-[(5-{[5-fluoro-2′-(propan-2-yl)[1,1′-biphenyl]-2-yl]oxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-olStep 1 benzyl{(1R,3S,4R)-3-[(5-{[5-fluoro-2′-(propan-2-yl)[1,1′-biphenyl]-2-yl]oxy}pyrimidin-4-yl)(methyl)amino]-4-hydroxycyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 2 ofReference Example C-25, using the compound obtained in Step 1 ofReference Example C-25 and 2-isopropylphenylboronic acid (CAS:89787-12-2).

MS (m/z): 571 (M+H)⁺.

Step 2(1R,2S,4R)-4-amino-2-[(5-{[5-fluoro-2′-(propan-2-yl)[1,1′-biphenyl]-2-yl]oxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 1.

MS (m/z): 437 (M+H)⁺.

Reference Example C-27(1R,2S,4R)-4-amino-2-[(5-{4-fluoro-2-[1-(propan-2-yl)-1H-pyrazol-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-olStep 1 benzyl{(1R,3S,4R)-3-[(5-{4-fluoro-2-[1-(propan-2-yl)-1H-pyrazol-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]-4-hydroxycyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 2 ofReference Example C-25, using the compound obtained in Step 1 ofReference Example C-25 and 1-isopropylpyrazole-5-boronic acid (CAS:839714-33-9).

¹H-NMR (CDCl₃) δ: 1.41-1.45 (6H, m), 1.63-1.66 (1H, m), 2.10-2.25 (3H,m), 3.06 (3H, s), 3.26-3.29 (1H, m), 4.00-4.03 (1H, m), 4.20-4.25 (1H,m), 4.32-4.38 (2H, m), 5.09 (2H, s), 5.29-5.31 (1H, m), 6.16-6.16 (1H,m), 6.74-6.77 (1H, m), 7.06-7.12 (2H, m), 7.33-7.36 (4H, m), 7.57 (1H,s), 7.85 (1H, s), 8.39 (1H, s).

MS (m/z): 561 (M+H)⁺.

Step 2(1R,2S,4R)-4-amino-2-[(5-{4-fluoro-2-[1-(propan-2-yl)-1H-pyrazol-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.41-1.59 (9H, m), 1.68-1.75 (2H, m), 1.86-1.93 (1H,m), 2.09-2.11 (1H, m), 3.12-3.14 (3H, m), 3.53-3.58 (1H, m), 4.26-4.27(1H, m), 4.35-4.38 (1H, m), 4.56-4.59 (1H, m), 6.18 (1H, d, J=1.8 Hz),6.76-6.78 (1H, m), 7.02-7.11 (2H, m), 7.58 (1H, d, J=1.8 Hz), 7.83 (1H,s), 8.40 (1H, s).

MS (m/z): 427 (M+H)⁺.

Reference Example C-286-[(4-{[(1S,2R,4R)-4-amino-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-2,3-difluoro-N,N-di(propan-2-yl)benzamideStep 1 benzyl{(1R,3S,4R)-3-[(5-{2-[di(propan-2-yl)carbamoyl]-3,4-difluorophenoxy}pyrimidin-4-yl)(methyl)amino]-4-hydroxycyclopentyl}carbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 14 ofReference Example A-1 and the compound obtained in Step 5 of ReferenceExample B-10.

¹H-NMR (CDCl₃) δ: 1.09-1.10 (1H, m), 1.20-1.25 (5H, m), 1.43-1.46 (3H,m), 1.53-1.55 (3H, m), 1.64-1.73 (1H, m), 2.08-2.25 (2H, m), 2.32-2.41(1H, m), 3.20-3.22 (3H, m), 3.49-3.54 (1H, m), 3.78-3.84 (1H, m),4.01-4.08 (1H, m), 4.23-4.47 (2H, m), 5.09 (2H, s), 5.30-5.32 (1H, m),6.38-6.42 (1H, m), 7.03-7.08 (1H, m), 7.30-7.38 (5H, m), 7.89 (1H, d,J=14.1 Hz), 8.44 (1H, d, J=3.7 Hz).

MS (m/z): 598 (M+H)⁺.

Step 26-[(4-{[(1S,2R,4R)-4-amino-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-2,3-difluoro-N,N-di(propan-2-yl)benzamide

The title compound was obtained in the same manner as in Step 2 ofReference Example C-14, using the compound obtained in the above Step 1.

MS (m/z): 464 (M+H)⁺.

Reference Example C-29 tert-butyl[(1S,3R)-3-{[6-(cyclopropylmethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]carbamate

To tert-butyl [(1S,3R)-3-(methylamino)cyclopentyl]carbamate (200 mg) andthe compound (210 mg) obtained in Step 2 of Reference Example B-2 wereadded 2-propanol (5 mL) and DIPEA (0.325 mL), and the mixture was heatedunder reflux for 6 hr. Ethyl acetate and saturated brine were added tothe reaction solution, and the mixture was subjected to liquidseparation. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (205 mg) as an oil.

¹H-NMR (CDCl₃) δ: 0.27-0.31 (2H, m), 0.60-0.64 (2H, m), 1.02-1.12 (1H,m), 1.46 (9H, s), 1.58-1.71 (2H, m), 1.84-2.13 (3H, m), 2.35-2.42 (1H,m), 2.76 (2H, d, J=7.3 Hz), 3.25 (3H, s), 3.97-4.04 (1H, m), 4.97-5.08(1H, m), 5.11-5.20 (1H, m), 7.12 (1H, s), 8.39 (1H, s).

MS (m/z): 403 (M+H)⁺.

Reference Example C-30(1R,2R,4S)-2-amino-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride Step 1 tert-butyl[(1R,2R,4S)-2-(methoxymethoxy)-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

A mixture of the compound (0.370 g) obtained in Step 7 of ReferenceExample A-12, 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(0.358 g), DIPEA (0.469 mL) and 2-propanol (15.0 mL) was stirred at 100°C. for 10 hr. The mixture was concentrated under reduced pressure, andthe obtained residue was subjected to silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (0.592 g) as asolid.

¹H-NMR (DMSO-D₆, 80° C.) δ: 1.40 (9H, s), 1.65-1.73 (1H, m), 1.85-1.92(1H, m), 2.04-2.11 (1H, m), 2.21-2.28 (1H, m), 3.21 (3H, s), 3.28 (3H,s), 3.75-3.83 (1H, m), 3.95-4.06 (3H, m), 4.60 (1H, d, J=6.7 Hz), 4.67(1H, d, J=6.7 Hz), 5.24-5.34 (1H, m), 6.73-6.82 (1H, m), 7.61 (1H, s),8.34 (1H, s).

Step 2(1R,2R,4S)-2-amino-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

A mixture of the compound (0.162 g) obtained in the above Step 1,hydrogen chloride (4 mol/L, 1,4-dioxane solution, 5 mL) and methanol (5mL) was stirred at room temperature for 1 hr. The mixture wasconcentrated, and the residue was dried to give the title compound(0.138 g) as a solid. This was directly used in the next step.

Reference Example C-31(1S,2R,4S)-2-amino-4-{methyll[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olStep 1 tert-butyl[(1R,2S,4S)-2-(methoxymethoxy)-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate

A mixture of tert-butyl[(1R,2S,4S)-2-(methoxymethoxy)-4-(methylamino)cyclopentyl]carbamate(0.414 g), 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine(0.400 g), 2-propanol (15.0 mL) and DIPEA (0.525 mL) was stirred at 100°C. for 4 hr. The mixture was concentrated under reduced pressure, andthe obtained residue was subjected to silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (0.662 g) as asolid.

¹H-NMR (CDCl₃) δ: 1.46 (9H, s), 1.75-1.90 (2H, m), 2.24-2.42 (2H, m),3.30 (3H, s), 3.42 (3H, s), 3.63 (2H, q, J=10.1 Hz), 3.93-4.03 (1H, m),4.07-4.14 (1H, m), 4.70 (1H, d, J=6.7 Hz), 4.76 (1H, d, J=6.7 Hz),5.11-5.19 (1H, m), 5.37-5.50 (1H, m), 7.33 (1H, s), 8.42 (1H, s).

Step 2(1S,2R,4S)-2-amino-4-{methyll[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.This compound was directly used in the next reaction.

Reference Example C-32(1R,3S,5R)-5-(1,3-oxazol-2-yl)-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine(racemate) Step 1 methyl(1S,3S,5R)-3-{[(benzyloxy)carbonyl]amino}-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxylate(racemate)

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 4 ofReference Example A-9 andchloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine producedaccording to the method described in a literature (cancer cell 2015, 27,589-602.).

¹H-NMR (CDCl₃) δ: 1.12-1.47 (3H, m), 2.31-2.53 (3H, m), 2.60-2.72 (1H,m), 3.53-3.87 (6H, m), 4.31-4.44 (1H, m), 4.73-4.83 (1H, m), 5.00-5.20(3H, m), 7.03 (1H, s), 7.29-7.40 (5H, m), 8.47 (1H, s).

MS (m/z): 523 (M+H)⁺.

Step 2(1S,3S,5R)-3-{[(benzyloxy)carbonyl]amino}-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxylicacid (racemate)

A mixture of the compound (1.00 g) obtained in the above Step 1, THF(15.0 mL) and lithium hydroxide monohydrate (210 mg) was stirred at roomtemperature for 2 hr. The reaction mixture was concentrated underreduced pressure, the residue was acidified (pH3-4) to with 2Nhydrochloric acid (2.00 mL) at 0° C., and the mixture was extracted withethyl acetate. The organic layer was washed with saturated brine, anddried over anhydrous sodium sulfate. The solvent was evaporated underreduced pressure to give the title compound (0.949 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.15-1.41 (3H, m), 1.96-2.17 (3H, m), 2.38-2.52 (1H,m), 3.43-3.59 (1H, m), 4.01-4.13 (2H, m), 4.14-4.27 (1H, m), 5.01 (2H,s), 7.27-7.44 (6H, m), 7.64 (1H, s), 7.86-7.92 (1H, m), 8.34 (1H, s).

MS (m/z): 509 (M+H)⁺.

Step 3 benzyl [(1S,3S,5R)-3-[(2,2-dimethoxyethyl)carbamoyl]-5-20{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate(racemate)

To a solution of the compound (0.455 g) obtained in the above Step 2 inDMF (4.50 mL) were successively added COMU (CAS: 1075198-30-9) (0.471g), DIPEA (0.234 mL) and aminoacetaldehyde dimethylacetal (CAS:22483-09-6) (0.118 mL) at 0° C., and the mixture was stirred at roomtemperature for 16 hr. The reaction mixture was poured into water, andextracted with a mixed solvent of ethyl acetate/diethyl ether. Theorganic layer was washed successively with water (three times) andsaturated brine, and dried over anhydrous sodium sulfate. The solventwas evaporated under reduced pressure, and the obtained residue wassubjected to silica gel column chromatography (n-hexane/ethyl acetate,followed by ethyl acetate/methanol) to give the title compound (0.491 g)as a solid.

¹H-NMR (CDCl₃) δ: 1.21-1.37 (1H, m), 1.45-1.64 (2H, m), 2.13-2.29 (2H,m), 2.38-2.50 (2H, m), 3.33-3.43 (8H, m), 3.56-3.68 (2H, m), 3.70-3.83(1H, m), 4.31-4.47 (2H, m), 4.94-5.10 (3H, m), 5.43-5.60 (1H, m),5.77-5.87 (1H, m), 7.10 (1H, br s), 7.28-7.38 (5H, m), 8.46 (1H, s).

MS (m/z): 596 (M+H)⁺.

Step 4 benzyl[(1S,3S,5R)-3-[(2-oxoethyl)carbamoyl]-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate(racemate)

To a solution of the compound (0.491 g) obtained in the above Step 3 indichloromethane (80.0 mL) was added trifluoroacetic acid (24.0 mL) atroom temperature, and the mixture was stirred at the same temperaturefor 16 hr. The mixture was stirred with heating under reflux foradditional 2 hr. The solvent was evaporated under reduced pressure,ethyl acetate and saturated aqueous sodium hydrogencarbonate solutionwere added to the obtained residue, and the mixture was extracted withethyl acetate. The organic layer was washed with saturated brine, anddried over anhydrous sodium sulfate. The solvent was evaporated underreduced pressure, and the residue was dried to give the title compoundas a solid. This was directly used in the next step.

MS (m/z): 550 (M+H)⁺.

Step 5 benzyl[(1S,3S,5R)-3-(1,3-oxazol-2-yl)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate(racemate)

To a solution of the compound (0.382 g) obtained in the above Step 4 indichloromethane (7.00 mL) were added triphenylphosphine (0.273 g),hexachloroethane (CAS: 67-72-1) (0.247 g) and TEA (0.193 mL) at roomtemperature, and the mixture was stirred for 3 days. The reactionmixture was concentrated under reduced pressure, and the residue wassubjected to silica gel column chromatography (ethyl acetate/methanol)to give a mixture containing the objective product as an oil. Thiscompound was used in the next step without further purification.

Step 6 (1R,3S,5R)-5-(1,3-oxazol-2-yl)-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine(racemate)

To a solution of the compound obtained in the above Step 5 inacetonitrile (15.0 mL) was added iodotrimethylsilane (0.239 mL) at 0°C., and the mixture was stirred at room temperature for 2 hr. Additionaliodotrimethylsilane (0.239 mL) was added thereto, and the mixture wasstirred for 0.5 hr. The solvent was evaporated under reduced pressure,and the obtained residue was subjected to amino silica gel columnchromatography (n-hexane/ethyl acetate, followed by ethylacetate/methanol) to give the title compound (0.0472 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.13-1.72 (5H, m), 2.29-2.44 (2H, m), 2.54-2.63 (1H,m), 3.05-3.18 (2H, m), 3.57-3.70 (2H, m), 4.37-4.50 (1H, m), 5.11-5.25(1H, m), 6.97-7.07 (2H, m), 7.57 (1H, s), 8.48 (1H, s).

MS (m/z): 398 (M+H)⁺.

Reference Example C-33 tert-butyl[(1R,2R,3S,4S)-2,3-dihydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate(isomer A) tert-butyl[(1R,2S,3R,4S)-2,3-dihydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate(isomer B) Step 1 tert-butyl prop-2-en-1-yl(1R,3S)-cyclopent-4-ene-1,3-diylbiscarbamate

The title compound was obtained in the same manner as in Step 4 ofReference Example A-6, using(1S,4R)-4-[(tert-butoxycarbonyl)amino]cyclopent-2-ene-1-carboxylic acid(1.00 g) synthesized according to the method described in a literature(Eur. J. Org. Chem. 2013, 17, 3477-3493).

¹H-NMR (CDCl₃) δ: 1.45-1.46 (11H, m), 2.81-2.90 (1H, m), 4.60-4.91 (5H,m), 5.21-5.23 (1H, m), 5.30-5.32 (1H, m), 5.79-5.96 (3H, m).

MS (m/z): 183 (M+H−Boc)⁺.

Step 2 tert-butyl [(1R,4S)-4-aminocyclopent-2-en-1-yl]carbamate

A mixture of the compound (2.87 g) obtained in the above Step 2,dichloromethane (15 mL), pyrrolidine (0.366 mL) andtetrakis(triphenylphosphine)palladium(0) (41 mg) was stirred at roomtemperature for 1 hr. The mixture was concentrated under reducedpressure, and the residue was purified by silica gel columnchromatography (ethyl acetate/methanol) to give the title compound (1.35g, containing a small amount of a stereoisomer) as an oil.

¹H-NMR (CDCl₃) δ: 1.11-1.18 (1H, m), 1.45 (9H, s), 2.76-2.83 (1H, m),3.85-3.88 (1H, m), 4.61-4.67 (2H, m), 5.77-5.85 (2H, m).

MS (m/z): 199 (M+H)⁺.

Step 3 tert-butyl{(1R,4S)-4-[(2-nitrobenzene-1-sulfonyl)amino]cyclopent-2-en-1-yl}carbamate

The title compound (containing a small amount of a stereoisomer) wasobtained in the same manner as in Step 12 of Reference Example A-1,using the compound obtained in the above Step 2.

1H-NMR (CDCl₃) δ: 1.37-1.40 (10H, m), 2.63-2.67 (1H, m), 4.46-4.49 (2H,m), 4.71-4.75 (1H, m), 5.76-5.83 (3H, m), 7.73-7.78 (2H, m), 7.85-7.89(1H, m), 8.17-8.18 (1H, m).

Step 4 tert-butyl{(1R,4S)-4-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopent-2-en-1-yl}carbamate

The title compound (containing a small amount of a stereoisomer) wasobtained in the same manner as in Step 13 of Reference Example A-1,using the compound obtained in the above Step 3.

¹H-NMR (CDCl₃) δ: 1.37-1.41 (10H, m), 2.71-2.74 (4H, m), 4.58-4.60 (2H,m), 4.97-4.99 (1H, m), 5.69-5.70 (1H, m), 5.89-5.90 (1H, m), 7.63-7.65(1H, m), 7.67-7.74 (2H, m), 8.02-8.07 (1H, m).

MS (m/z): 298 (M+H)⁺.

Step 5 tert-butyl [(1R,4S)-4-(methylamino)cyclopent-2-en-1-yl]carbamate

A mixture of the compound (887 mg) obtained in the above Step 4,4-tert-butylbenzenethiol (0.751 mL), potassium carbonate (1.23 g) andDMF (11.2 mL) was stirred at 40° C. for 2.5 hr. Water was added to thereaction solution, and the mixture was extracted with ethyl acetate. Theorganic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (ethyl acetate/methanol) togive the title compound (498 mg, containing a small amount of astereoisomer) as an oil.

¹H-NMR (CDCl₃) δ: 1.23-1.31 (1H, m), 1.45 (9H, s), 2.41-2.44 (3H, m),2.66-2.74 (1H, m), 3.61-3.64 (1H, m), 4.64-4.75 (2H, m), 5.82-5.92 (2H,m).

MS (m/z): 213 (M+H)⁺.

Step 6 tert-butyl[(1R,4S)-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopent-2-en-1-yl]carbamate

To a mixture of the compound (622 mg) obtained in the above Step 5 and2-propanol (23.4 mL) was added DIPEA (0.816 mL), and the mixture wasstirred overnight at 90° C. The mixture was concentrated under reducedpressure, and the residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give a solid (800 mg). 750 mgof the obtained solid was purified by chiral column (DAICEL, CHIRALPAK(registered trademark, Daicel Corporation) IA, n-hexane/2-propanol) togive the title compound (541 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.45-1.48 (10H, m), 2.96-2.99 (1H, m), 3.19 (3H, s),3.64 (2H, q, J=10.0 Hz), 4.70-4.72 (2H, m), 5.85-5.86 (1H, m), 5.93-5.98(2H, m), 7.36 (1H, s), 8.44 (1H, s).

MS (m/z): 429 (M+H)⁺.

Step 7 tert-butyl[(1R,2R,3S,4S)-2,3-dihydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate(isomer A) tert-butyl[(1R,2S,3R,4S)-2,3-dihydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]carbamate(isomer B)

A mixture of the compound (172 mg) obtained in the above Step 6, osmiumtetraoxide (4% aqueous solution) (0.051 mL), 4-methylmorpholine N-oxide(70.5 mg), acetone (3.6 mL) and water (0.40 mL) was stirred at roomtemperature for 4 hr. Additional 4-methylmorpholine N-oxide (70.5 mg)and osmium tetraoxide (4% aqueous solution) (0.13 mL) were added to thereaction solution, and the mixture was stirred for additional 3 hr.Aqueous sodium thiosulfate solution was added to the reaction solution,and the mixture was extracted with ethyl acetate. The organic layer waswashed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (isomer A, an earlier eluted component) (44.9 mg), and thetitle compound (isomer B, a later eluted component) (56.4 mg),respectively as a solid.

the isomer A (earlier eluted component)

¹H-NMR (CD₃OD) δ: 1.46 (9H, s), 2.16-2.18 (2H, m), 3.47 (3H, s),3.86-3.89 (3H, m), 4.04-4.05 (1H, m), 4.36-4.38 (1H, m), 5.23-5.27 (1H,m), 7.65 (1H, s), 8.30 (1H, s).

MS (m/z): 463 (M+H)⁺.

the isomer B (later eluted component)

¹H-NMR (CD₃OD) δ: 1.46 (9H, s), 1.60-1.67 (1H, m), 2.42 (1H, dt, J=15.5,6.7 Hz), 3.33 (3H, s), 3.80 (1H, td, J=7.7, 3.7 Hz), 3.85-3.93 (3H, m),4.30 (1H, dd, J=8.6, 5.5 Hz), 5.04 (1H, q, J=9.6 Hz), 7.67 (1H, s), 8.32(1H, s).

MS (m/z): 463 (M+H)⁺.

Reference Example D-1 5-formyl-1H-indole-2-carbonitrile

Sodium hydride (purity 55%, 628 mg) was placed in a reaction container,the container was subjected to nitrogen substitution, and THF (30 mL)was added thereto while ice-cooling. A THF solution (42 mL) of5-bromo-1H-indole-2-carbonitrile (1.59 g) synthesized according to themethod described in a literature (WO 2014/164749) was added dropwisethereto, and the mixture was stirred at room temperature for 15 min. Thereaction solution was cooled to −78° C., tert-butyllithium (1.65 mol/L,n-pentane solution, 10.9 mL) was added dropwise thereto over 20 min, andthe mixture was stirred at the same temperature for 45 min. DMF (2.8 mL)was added dropwise thereto over 5 min, and the mixture was stirred at−78° C. for 45 min. Acetic acid (4.1 mL) was added dropwise thereto, andthe mixture was allowed to warm to room temperature. Ethyl acetate and0.5N aqueous hydrochloric acid solution were added to the reactionsolution. The insoluble substance was removed by filtration, and thefiltrate was subjected to liquid separation. The aqueous layer wasextracted with ethyl acetate, and the organic layers were combined,washed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (946 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 7.61-7.65 (2H, m), 7.84 (1H, d, J=9.1 Hz), 8.35 (1H,s), 10.02 (1H, s), 12.90 (1H, s).

Reference Example D-25-formyl-4-methyl-1-{[1-(triphenylmethyl)-1H-pyrazol-4-yl]methyl}-1H-indole-2-carbonitrileStep 1 5-formyl-4-methyl-1H-indole-2-carbonitrile

The title compound was obtained in the same manner as in ReferenceExample D-1, using 5-bromo-4-methyl-1H-indole-2-carbonitrile (1.22 g)synthesized according to the method described in a literature (cancercell 2015, 27, 589-602.).

¹H-NMR (CD₃CN) δ: 2.85 (3H, s), 7.42 (1H, d, J=8.5 Hz), 7.50 (1H, s),7.81 (1H, d, J=8.5 Hz), 10.37 (1H, s).

Step 25-formyl-4-methyl-1-{[1-(triphenylmethyl)-1H-pyrazol-4-yl]methyl}-1H-indole-2-carbonitrile

Under argon atmosphere, to a mixture of the compound (0.250 g) obtainedin the above Step 1, cesium carbonate (0.619 g) and DMF (50 mL) wasadded a mixture of 4-(bromomethyl)-1-(triphenylmethyl)-1H-pyrazole(0.821 g) synthesized according to the method described in a literature(J. Med. Chem. 2016, 59(3), 892-913.) and DMF (90 mL) at 0° C., and themixture was stirred at room temperature for 1 hr. Water was added to thereaction solution, and the mixture was extracted with ethylacetate/diethyl ether. The solid of the organic layer was collected byfiltration to give the title compound (0.369 g) as a solid. The filtratewas concentrated, diethyl ether was added to the residue, and the solidwas collected by filtration to give the title compound (0.193 g) as asolid.

¹H-NMR (CDCl₃) δ: 2.88 (3H, s), 5.32 (2H, s), 7.07-7.09 (6H, m),7.26-7.31 (10H, m), 7.37 (1H, s), 7.44 (1H, s), 7.52 (1H, s), 7.88 (1H,d, J=9.1 Hz), 10.42 (1H, s).

Reference Example D-34-bromo-1-(methanesulfonyl)-1H-pyrrolo[2,3-c]pyridine

To a mixture of 4-bromo-1H-pyrrolo[2,3-c]pyridine (CAS: 69872-17-9) (401mg), DIPEA (1.05 mL) and dichloromethane (15 mL) was added dropwisemethanesulfonyl chloride (0.281 mL) under ice-cooling. The mixture wasstirred at room temperature for 10 min, saturated aqueous sodiumhydrogencarbonate solution was added to the reaction solution underice-cooling, and the mixture was extracted with ethyl acetate. Theorganic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure, and the residuewas subjected to silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (394 mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.24 (3H, s), 6.82 (1H, d, J=3.7 Hz), 7.66 (1H, d,J=3.7 Hz), 8.59 (1H, s), 9.17 (1H, s).

MS (m/z): 275, 277 (M+H)⁺.

Reference Example D-4 4-bromo-1-methyl-1H-pyrazolo[3,4-c]pyridine

To a mixture of 4-bromo-1H-pyrazolo[3,4-c]pyridine (CAS: 1032943-43-3)(262 mg), cesium carbonate (863 mg) and DMF (6.5 mL) was added dropwisemethyl iodide (CAS: 74-88-4) (0.107 mL) under ice-cooling. The mixturewas stirred at room temperature for 20 hr, water was added to thereaction solution under ice-cooling, and the mixture was extracted withethyl acetate. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure,and the residue was subjected to silica gel column chromatography(chloroform/methanol) to give the title compound (160 mg) as an oil.

¹H-NMR (CDCl₃) δ: 4.21 (3H, s), 8.06 (1H, s), 8.40 (1H, s), 8.89 (1H,s).

MS (m/z): 212, 214 (M+H)⁺.

Reference Example D-5 4-bromo-1-(methanesulfonyl)-1H-indazole

To a suspension of sodium hydride (CAS: 7646-69-7) (purity 55%, 734 mg)and DMF (100 mL) was added 4-bromoindazole (3.00 g) little by littleunder ice-cooling, the mixture was stirred at room temperature for 15min, and methanesulfonyl chloride (2.27 g) was added dropwise theretounder ice-cooling. The mixture was stirred at room temperature for 3 hr,and the reaction solution was weakly acidified with 1N hydrochloric acidunder ice-cooling, and extracted with ethyl acetate. The organic layerwas washed with saturated brine, dried over anhydrous sodium sulfate,and concentrated under reduced pressure, and the residue was subjectedto silica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (2.93 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.30 (3H, s), 7.43 (1H, dd, J=8.6, 7.4 Hz), 7.53 (1H,d, J=8.6 Hz), 8.04 (1H, d, J=7.4 Hz), 8.32 (1H, s).

MS (m/z): 275, 277 (M+H)⁺.

Reference Example D-6 4-bromo-6-fluoro-1-(methanesulfonyl)-1H-indazole

The title compound was obtained in the same manner as in ReferenceExample D-5, using 4-bromo-6-fluoro-1H-indazole (CAS: 885520-35-4).

¹H-NMR (CDCl₃) δ: 3.32 (3H, s), 7.35 (1H, dd, J=8.6, 1.8 Hz), 7.76 (1H,dd, J=8.3, 1.8 Hz), 8.27 (1H, d, J=1.2 Hz).

Reference Example D-7 N-(3-bromophenyl)-N-methylmethanesulfonamide

The title compound was obtained in the same manner as in ReferenceExample D-3, using 3-bromo-N-methyl-aniline.

¹H-NMR (DMSO-D₆) δ: 2.98 (3H, s), 3.24 (3H, s), 7.33-7.46 (2H, m),7.49-7.54 (1H, m), 7.61-7.65 (1H, m).

MS (m/z): 264, 266(M+H)⁺.

Reference Example D-8 1-bromo-2,4-difluoro-3-methoxybenzene

The title compound was obtained in the same manner as in ReferenceExample D-4, using 3-bromo-2,6-difluoro-phenol.

¹H-NMR (CDCl₃) δ: 4.01 (3H, s), 6.82-6.85 (1H, m), 7.19-7.20 (1H, m).

Reference Example D-9 (5-bromo-3-methoxypyridin-2-yl)methanol

A mixture of 5-bromo-3-methoxypyridine-2-carbaldehyde (1.22 g), sodiumborohydride (0.214 g) and ethanol (20.0 mL) was stirred for 1 hr underice-cooling. 1N Hydrochloric acid was added thereto, and the mixture wasextracted with ethyl acetate. The organic layers were combined, washedwith saturated brine, dried over anhydrous magnesium sulfate, andconcentrated under reduced pressure. The obtained residue was subjectedto silica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (0.819 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.87 (3H, s), 3.94 (1H, t, J=4.9 Hz), 4.69 (2H, d,J=4.9 Hz), 7.28 (1H, d, J=1.5 Hz), 8.23 (1H, d, J=1.5 Hz).

Reference Example D-10 5-bromo-3-methoxy-N-methylpyridin-2-amine

To a mixture of 5-bromo-3-methoxy-pyridin-2-amine (1.00 g) and THF (25.0mL) was sodium hydride (purity 55%, 0.431g) at 0° C., and the mixturewas stirred for 10 min. Methyl iodide (0.370 mL) was added thereto, andthe mixture was stirred at the same temperature for 1 hr. Ice was addedto the reaction solution, and the reaction mixture was extracted withdiethyl ether. The organic layer was washed successively with water andsaturated brine, dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure. The obtained residue was subjectedto silica gel column chromatography (n-hexane/ethyl acetate) to give thetitle compound (0.960 g) as a solid.

¹H-NMR (CDCl₃) δ: 2.96-3.01 (3H, m), 3.82 (3H, s), 4.85-4.98 (1H, m),6.89 (1H, d, J=1.8 Hz), 7.78 (1H, d, J=1.8 Hz).

MS (m/z): 217, 219 (M+H)⁺.

Reference Example D-114-[6-(dimethylamino)-5-methoxypyridin-3-yl]benzaldehyde Step 15-bromo-3-methoxy-N,N-dimethylpyridin-2-amine

To a mixture of 5-bromo-3-methoxypyridin-2-amine (3.00 g) and THF (75mL) was added sodium hydride (purity 55%, 2.58 g) under ice-cooling.After 10 min, methyl iodide (2.77 mL) was added thereto, and the mixturewas stirred at same temperature for 1 hr, and then at room temperaturefor 16 hr. Water was added to the reaction solution, and the mixture wasextracted with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (3.41g) as an oil.

¹H-NMR (CDCl₃) δ: 2.97 (6H, s), 3.83 (3H, s), 7.07 (1H, s), 7.86 (1H,s).

MS (m/z): 231, 233 (M+H)⁺.

Step 2 4-[6-(dimethylamino)-5-methoxypyridin-3-yl]benzaldehyde

A mixture of 5-bromo-3-methoxy-N,N-dimethylpyridin-2-amine (3.41 g),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (3.49 g),tetrakis(triphenylphosphine)palladium(0) (1.71 g), sodium carbonate(6.26 g), water (62 mL) and 1,4-dioxane (184 mL) was stirred undernitrogen atmosphere at 100° C. for 3 hr. Water was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate).The obtained residue was suspended in n-hexane/ethyl acetate, and thesolid was collected by filtration to give the title compound (2.37 g) asa solid.

¹H-NMR (CDCl₃) δ: 3.08 (6H, s), 3.94 (3H, s), 7.23 (1H, d, J=1.8 Hz),7.71 (2H, d, J=8.6 Hz), 7.94 (2H, d, J=8.6 Hz), 8.16 (1H, d, J=1.8 Hz),10.04 (1H, s).

MS (m/z): 257 (M+H)⁺.

Reference Example D-12 5-bromo-3-methoxy-N,N-bis[(²H₃)methyl]pyridin-2-amine

The title compound was obtained in the same manner as in Step 1 ofReference Example D-11 except that methyl iodide-D3 (CAS: 865-50-9) wasused instead of iodomethane.

¹H-NMR (CDCl₃) δ: 3.84 (3H, s), 7.08 (1H, d, J=1.8 Hz), 7.87 (1H, d,J=1.8 Hz).

MS (m/z): 237, 239 (M+H)⁺.

Reference Example D-13 6-chloro-4-methoxy-N,N-dimethylpyridazin-3-amine

A mixture of 3,6-dichloro-4-methoxypyridazine (CAS: 70952-62-4) (0.506g) synthesized according to the method described in a literature (J.Org. Chem. 2014, 79, 10311-10322), THF (2.1 mL) and dimethylamine (CAS:124-40-3) (concentration 2.0 mol/L, THF solution, 2.1 mL) was stirred atroom temperature for 21 hr. Then, additional dimethylamine (CAS:124-40-3) (concentration 2.0 mol/L, THF solution, 6.4 mL) was addedthereto, and the mixture was stirred at room temperature for 21 hr.Then, additional dimethylamine (CAS: 124-40-3) (concentration 2.0 mol/L,THF solution, 8.4 mL) was added thereto, and the mixture was stirred atroom temperature for 24 hr. The reaction mixture was concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound(0.110 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.08 (6H, s), 3.92 (3H, s), 6.65 (1H, s).

Reference Example D-14 6-chloro-4-methoxy-3-(2-methoxyethoxy)pyridazine

To a suspension of sodium hydride (purity 55%, 203 mg) and toluene (40mL) was added 2-methoxyethanol (0.330 mL) little by little underice-cooling, and the mixture was stirred at room temperature for 15 min,and 3,6-dichloro-4-methoxy-pyridazine (750 mg) was added thereto littleby little under ice-cooling. The mixture was stirred at room temperaturefor 4 hr, and the reaction solution was weakly acidified with 1Nhydrochloric acid under ice-cooling, and extracted with ethyl acetate.The organic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure, and the residuewas subjected to silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (295 mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.44 (3H, s), 3.82 (2H, t, J=4.9 Hz), 3.93 (3H, s),4.68 (2H, t, J=4.9 Hz), 6.76 (1H, s).

MS (m/z): 219 (M+H)⁺.

Reference Example D-156-chloro-3-{[1-(methanesulfonyl)piperidin-4-yl]oxy}-4-methoxypyridazine

To a solution of sodium hydride (purity 55%, 156 mg) in toluene (10 mL)was added 1-(methylsulfonyl)piperidin-4-ol (561 mg) under ice-cooling,and the mixture was stirred for 15 min.3,6-Dichloro-4-methoxy-pyridazine (400 mg) was added thereto at the sametemperature, and the mixture was stirred at 80° C. for 2.5 hr. Water wasadded to the reaction solution, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated brine, dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (288 mg) as a solid.

¹H-NMR (CDCl₃) δ: 2.01-2.04 (2H, m), 2.18-2.22 (2H, m), 2.83 (3H, s),3.18-3.24 (2H, m), 3.54-3.58 (2H, m), 3.94 (3H, s), 5.43-5.47 (1H, m),6.78 (1H, s).

MS (m/z): 322, 324 (M+H)⁺.

Reference Example D-164-{5-methoxy-6-[(pyridin-3-yl)oxy]pyridazin-3-yl}benzaldehyde Step 16-chloro-4-methoxy-3-[(pyridin-3-yl)oxy]pyridazine,3-chloro-4-methoxy-6-[(pyridin-3-yl)oxy]pyridazine

A mixture of 3,6-dichloro-4-methoxypyridazine (300 mg),3-hydroxypyridine (159 mg), DMF (11.2 mL) and potassium carbonate (463mg) was stirred at 120° C. for 2 hr. Water was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate) togive a mixture (88.6 mg) of the title two isomers as a solid. This wasdirectly used in the next step.

Step 2 4-{5-methoxy-6-[(pyridin-3-yl)oxy]pyridazin-3-yl}benzaldehyde

The title compound was obtained in the same manner as in Step 2 ofReference Example D-11, using the compound obtained in the above Step 1.

MS (m/z): 308 (M+H)⁺.

Reference Example D-176-chloro-4-methoxy-3-[(oxan-3-yl)oxy]pyridazine(racemate)

The title compound was obtained in the same manner as in ReferenceExample D-15 except that 3-hydroxytetrahydropyran was used instead of1-(methylsulfonyl)piperidin-4-ol.

¹H-NMR (CDCl₃) δ: 1.90-1.98 (3H, m), 2.13-2.17 (1H, m), 3.62-3.90 (4H,m), 3.92 (3H, s), 5.31-5.36 (1H, m), 6.76 (1H, s).

MS (m/z): 245, 247 (M+H)⁺.

Reference Example D-18 3-(azetidin-1-yl)-6-chloro-4-methoxypyridazine

A mixture of 3,6-dichloro-4-methoxy-pyridazine (4.00 g), azetidine (4.98g), THF (40 mL) and DIPEA (2.43 mL) was stirred at 0° C. for 1 hr, andthen at room temperature for 16 hr. Water was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayers were combined, washed with saturated brine, dried over anhydrousmagnesium sulfate, and concentrated under reduced pressure. The obtainedresidue was subjected to silica gel column chromatography to give thetitle compound (0.498 g) as a solid.

¹H-NMR (CDCl₃) δ: 2.38 (2H, quint., J=7.4 Hz), 3.83 (3H, s), 4.24 (4H,t, J=7.4 Hz), 6.53 (1H, s).

Reference Example D-19 6-chloro-4-methoxypyridazin-3-ol

A mixture of 3,6-dichloro-4-methoxypyridazine (2.00 g) and acetic acid(40 mL) was stirred at 110° C. for 3.5 hr. The reaction solution wasconcentrated under reduced pressure, the residue was suspended indichloromethane, and the insoluble substance was removed by filtration.The filtrate was concentrated, and the residue was subjected to silicagel column chromatography (dichloromethane/methanol) to give the titlecompound (0.572 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.85 (3H, s), 6.91 (1H, s), 13.04 (1H, br s).

MS (m/z): 161, 163 (M+H)⁺.

Reference Example D-20 6-chloro-5-methoxypyridazin-3-ol

To a mixture of 3,6-dichloro-4-methoxy-pyridazine (2.00 g) and potassiumacetate (1.21 g) were added acetic acid (29 mL)/water (5.8 mL), and themixture was stirred with heating in a microwave reactor at 140° C. for 1hr. After the completion of the reaction, water was added to thereaction solution, and the insoluble solid was collected by filtrationto give the title compound (739 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.87 (3H, s), 6.38 (1H, s), 12.83 (1H, s).

MS (m/z): 161, 163 (M+H)⁺.

Reference Example D-21 6-chloro-3-(difluoromethoxy)-4-methoxypyridazine

To a mixture of the compound (0.570 g) obtained in Reference ExampleD-19, 50% aqueous potassium hydroxide solution (3.5 mL), water (3.5 mL)and acetonitrile (7.0 mL) was added difluoromethyltrifluoromethanesulfonate (CAS: 1885-46-7) (1.35 mL) at 0° C., and themixture was stirred at room temperature for 4 hr. Difluoromethyltrifluoromethanesulfonate (1.35 mL) was added thereto at 0° C., and themixture was stirred at room temperature for 1 hr, and then at 60° C. for1 hr. Water was added to the reaction solution, and the mixture wasextracted with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was subjected to silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound(0.130 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.99 (3H, s), 6.94 (1H, s), 7.60 (1H, t, J=71.7 Hz).

MS (m/z): 211, 213 (M+H)⁺.

Reference Example D-22 3-chloro-6-(difluoromethoxy)-4-methoxypyridazine(isomer A) 6-chloro-2-(difluoromethyl)-5-methoxypyridazin-3(2H)-one(isomer B)

To a mixture of the compound (200 mg) obtained in Reference Example D-20and acetonitrile (2.5 mL) were added 50% aqueous potassium hydroxidesolution (1.25 mL) and water (1.25 mL). Difluoromethyltrifluoromethanesulfonate (0.475 mL) was added thereto underice-cooling, and the mixture was stirred at room temperature for 1 hr.Water was added to the reaction solution, and the mixture was extractedwith ethyl acetate. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (isomer A, anearlier eluted component, 139 mg), and the title compound (isomer B, alater eluted component, 46.5 mg), respectively as a solid.

the isomer A ¹H-NMR (CDCl₃) δ: 4.01 (3H, s), 6.52 (1H, s), 7.62 (1H, t,J=71.8 Hz).

MS (m/z): 211, 213 (M+H)⁺.

the isomer B ¹H-NMR (CDCl₃) δ: 3.94 (3H, s), 6.12 (1H, s), 7.59 (1H, t,J=58.3 Hz).

MS (m/z): 211, 213 (M+H)⁺.

The structural formulas of the isomer A and isomer B are estimatedstructures, and the structural formula of the isomer A and thestructural formula of the isomer B may be interchanged. The same appliesto the subsequent steps (Reference Example D-59 and Reference ExampleD-60, and Examples 110 and 111).

Reference Example D-23 6-chloro-3,4-dimethoxypyridazine

3,4,6-Trichloropyridazine (CAS: 6082-66-2) (2.57 g) was dissolved inmethanol (50 mL), and sodium methoxide (CAS: 124-41-4) (1.56 g) wasadded thereto. The mixture was stirred at 0° C. for 10 min, allowed towarm to room temperature, and stirred for 20 hr. The reaction solutionwas concentrated under reduced pressure, ethyl acetate and saturatedaqueous ammonium chloride solution were added thereto, and the mixturewas subjected to liquid separation operation. The organic layer waswashed with saturated brine, dried over anhydrous sodium sulfate,filtered, and concentrated under reduced pressure. The residue wassubjected to silica gel column chromatography (dichloromethane/ethylacetate), and the obtained solid was suspended in n-hexane/ethylacetate. The insoluble substance was collected by filtration, and driedto give the title compound (1.10 g) as a solid.

¹H-NMR (CDCl₃) δ: 3.95 (3H, s), 4.16 (3H, s), 6.77 (1H, s).

MS: m/z 175, 177 (M+H)⁺.

Reference Example D-24 6-chloro-3,4-bis [(²H₃)methyloxy] pyridazine

The title compound was obtained in the same manner as in ReferenceExample D-23 except that methanol-D3 was used instead of methanol.

¹H-NMR (CDCl₃) δ: 6.77 (1H, s).

MS (m/z): 181, 183 (M+H)⁺.

Reference Example D-255-(4-formylphenyl)-3-methoxypyridine-2-carbonitrile Step 13-fluoro-5-(4-formylphenyl)pyridine-2-carbonitrile

A mixture of 5-bromo-3-fluoro-pyridine-2-carbonitrile (CAS: 886373-28-0)(2.00 g), (4-formylphenyl)boronic acid (1.49 g),tetrakis(triphenylphosphine)palladium(0) (1.15 g), sodium carbonate(3.16 g), water (8.0 mL) and 1,4-dioxane (24 mL) was stirred at 100° C.for 4 hr. The reaction solution was diluted with ethyl acetate, washedsuccessively with water and saturated brine, dried over anhydrousmagnesium sulfate, and concentrated under reduced pressure. The obtainedresidue was subjected to silica gel column chromatography(n-hexane/ethyl acetate). The fraction was concentrated, and theobtained solid was subjected to slurry washing with n-hexane to give thetitle compound (1.28 g) as a solid.

¹H-NMR (CDCl₃) δ: 7.79 (2H, d, J=8.6 Hz), 7.83 (1H, dd, J=9.2, 1.8 Hz),8.07 (2H, d, J=8.6 Hz), 8.82-8.83 (1H, m), 10.12 (1H, s).

Step 2 5-(4-formylphenyl)-3-methoxypyridine-2-carbonitrile

A mixture of the compound (0.250 g) obtained in the above Step 1,potassium carbonate (0.458 g) and methanol (10.0 mL) was stirred at 70°C. for 10 min. The reaction solution was diluted with ethyl acetate,washed with water and saturated brine, dried over anhydrous magnesiumsulfate, and concentrated under reduced pressure. The obtained residuewas subjected to silica gel column chromatography (dichloromethane/ethylacetate) to give the title compound (0.163 g) as a solid.

¹H-NMR (CDCl₃) δ: 4.07 (3H, s), 7.51 (1H, d, J=1.8 Hz), 7.78 (2H, d,J=8.6 Hz), 8.05 (2H, d, J=8.6 Hz), 8.55 (1H, d, J=1.8 Hz), 10.11 (1H,s).

Reference Example D-26 4-(5,6-dimethoxypyridazin-3-yl)benzaldehyde

A mixture of the compound (1.10 g) obtained in Reference Example D-23,4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (CAS:128376-64-7) (2.17 g), tetrakis(triphenylphosphine)palladium(0) (CAS:14221-01-3) (0.720 g), sodium carbonate (1.70 g), 1,2-dimethoxyethane(30 mL) and water (10 mL) was stirred at 100° C. for 5 hr. The reactionsolution was allowed to cool to room temperature, saturated brine wasadded thereto, and the mixture was extracted with ethyl acetate. Theorganic layer was dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure. The residue was subjected to silicagel column chromatography (n-hexane/ethyl acetate, followed by ethylacetate/methanol), and the obtained solid was suspended inn-hexane/ethyl acetate. The resulting solid was collected by filtration,and dried to give the title compound (1.21 g) as a solid.

¹H-NMR (CDCl₃) δ: 4.05 (3H, s), 4.26 (3H, s), 7.20 (1H, s), 7.99-8.04(2H, m), 8.15-8.20 (2H, m), 10.10 (1H, s).

Reference Example D-27 tert-butyl4-(4-formylphenyl)-1H-pyrazole-1-carboxylate

A mixture of1-boc-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (1.43 g),4-bromobenzaldehyde (150 mg), cesium carbonate (3.17 g),[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloridedichloromethane adduct (397 mg), 1,4-dioxane (25.6 mL) and water (2.56mL) was stirred at 70° C. for 4 hr. Water was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (n-hexane/ethyl acetate) togive the title compound (842 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.69 (9H, s), 7.70 (2H, d, J=5.5 Hz), 7.92 (2H, d,J=5.5 Hz), 8.07 (1H, s), 8.43 (1H, s), 10.02 (1H, s).

MS (m/z): 173 (M−Boc+H)⁺.

The products described in the following Table 1-1 to Table 1-12 wereproduced from Raw Material 1 and Raw Material 2 described in the tables,in the same manner as in Reference Example D-26 or D-27.

TABLE 1-1 Ref. Ex. Product Raw Material 1 Raw Material 2 No. CompoundName, NMR, MS D-28

4-(6-chloro-5-methoxypyridazin-3-yl)benzaldehyde ¹H-NMR (CDCl₃) δ: 4.11(3H, s), 7.29 (1H, s), 8.01-8.07 (2H, m), 8.17-8.24 (2H, m), 10.12 (1H,s). MS (m/z): 249, 251 (M + H)⁺. D-29

4-[6-(dimethylamino)-5-methoxypyridazin-3-yl]benzaldehyde ¹H-NMR (CDCl₃)δ: 3.20 (6H, s), 4.01 (3H, s), 7.09 (1H, s), 7.99 (2H, d, J = 8.6 Hz),8.20 (2H, d, J = 8.6 Hz), 10.08 (1H, s). MS (m/z): 258 (M + H)⁺. D-30

4-(6-{bis[(²H₃)methyl]amino}-5-methoxypyridin-3-yl)benzaldehyde ¹H-NMR(CDCl₃) δ: 3.93 (3H, s), 7.23 (1H, d, J = 2.6 Hz), 7.70 (2H, d, J = 8.6Hz), 7.94 (2H, d, J = 8.6 Hz), 8.16 (1H, d, J = 2.5 Hz), 10.04 (1H, s).MS (m/z): 263 (M + H)⁺. D-31

4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}benzaldehyde ¹H-NMR (CDCl₃) δ:7.20 (1H, s), 8.01 (2H, d, J = 8.5 Hz), 8.18 (2H, d, J = 8.5 Hz), 10.10(1H, s). MS (m/z): 251 (M + H)⁺.

TABLE 1-2 D-32

4-(1,3-oxazol-2-yl)benzaldehyde ¹H-NMR (DMSO-D₆) δ: 7.50 (1H, s), 8.07(2H, d, J = 8.2 Hz), 8.20 (2H, d, J = 8.2 Hz), 8.35 (1H, s), 10.08 (1H,s). MS (m/z): 174 (M + H)⁺. D-33

2′,4′-difluoro-3′-methoxy[1,1′-biphenyl]-4-carbaldehyde ¹H-NMR (CDCl₃)δ: 4.05 (3H, s), 6.98-7.03 (1H, m), 7.09-7.11 (1H, m), 7.66-7.68 (2H,m), 7.96- 7.97 (2H, m), 10.07 (1H, s). MS (m/z): 249 (M + H)⁺. D-34

4-[5-(difluoromethoxy)-6-methylpyridin-3-yl]benzaldehyde ¹H-NMR (CDCl₃)δ: 2.60 (3H, s), 6.63 (1H, t, J = 72.6 Hz), 7.67 (1H, s), 7.73 (2H, d, J= 7.9 Hz), 7.99 (2H, d, J = 8.5 Hz), 8.64 (1H, s), 10.07 (1H, s). MS(m/z): 264 (M + H)⁺. D-35

ethyl 4-(5,6-dimethoxypyridazin-3-yl)cyclohex-3-ene-1-carboxylate(racemate) ¹H-NMR (CDCl₃) δ: 1.37 (3H, t, J = 7.0 Hz), 1.79-1.90 (1H,m), 2.18-2.26 (1H, m), 6.90 (1H, s), 2.51-2.56 (2H, m), 2.58-2.69 (2H,m), 2.85-2.94 (1H, m), 3.95 (3H, s), 4.16-4.22 (5H, m), 6.43- 6.47 (1H,m). MS (m/z): 293 (M + H)⁺.

TABLE 1-3 D-36

4-(5-methoxy-6-methylpyridin-3-yl)benzaldehyde ¹H-NMR (CDCl₃) δ: 2.53(3H, s), 3.93 (3H, s), 7.28 (1H, d, J = 1.8 Hz), 7.73-7.78 (2H, m),7.96-8.01 (2H, m), 8.35 (1H, d, J = 1.8 Hz), 10.08 (1H, s). D-37

3′,4′-difIuoro-5′-methoxy[1,1′-biphenyl]-4-carbaldehyde ¹H-NMR (CDCl₃)δ: 3.99 (3H, s), 6.97-7.07 (2H, m), 7.69 (2H, d, J = 8.3 Hz), 7.96 (2H,d, J = 8.3 Hz), 10.07 (1H, s). MS (m/z): 249 (M + H)⁺. D-38

4-(1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)benzaldehyde ¹H-NMR (CDCl₃)δ: 4.27 (3H, s), 7.90 (2H, d, J = 8.6 Hz), 8.07 (2H, d, J = 8.6 Hz),8.19 (1H, s), 8.48 (1H, s), 9.02 (1H, s), 10.13 (1H, s). MS (m/z): 238(M + H )⁺. D-39

4-(6-chloro-5-methoxypyridin-3-yl)benzaldehyde ¹H-NMR (DMSO-D₆) δ: 4.02(3H, s), 7.39 (1H, d, J = 1.8 Hz), 7.74 (2H, d, J = 8.0 Hz), 8.01 (2H,d, J = 8.0 Hz), 8.26 (1H, d, J = 1.8 Hz), 10.09 (1H, s). MS (m/z): 248,250 (M + H)⁺.

TABLE 1-4 D-40

4-[1-(methanesulfonyl)-1H-indazol-4-yl]benzaldehyde ¹H-NMR (CDCl₃) δ:3.32 (3H, s), 7.49 (1H, d, J = 6.7 Hz), 7.68 (1H, dd, J = 8.0, 6.7 Hz),7.80 (2H, d, J = 8.6 Hz), 8.07 (2H, d, J = 8.6 Hz), 8.16 (1H, d, J = 8.0Hz), 8.39 (1H, s), 10.13 (1H, s). MS (m/z): 301 (M + H)⁺. D-41

4-[6-fluoro-1-(methanesulfonyl)-1H-indazol-4-yl]benzaldehyde ¹H-NMR(CDCl₃) δ: 3.34 (3H, s), 7.26 (1H, dd, J = 9.2, 1.8 Hz), 7.78 (2H, d, J= 8.6 Hz), 7.84- 7.87 (1H, m), 8.08 (2H, d, J = 8.6 Hz), 8.33 (1H, d, J= 1.2 Hz), 10.13 (1H, s). MS (m/z): 319 (M + H)⁺. D-42

4-(2,3-dihydro[1,4]dioxino[2,3-b]pyridin-7-yl)benzaldehyde ¹H-NMR(CDCl₃) δ: 4.30-4.33 (2H, m), 4.46-4.48 (2H, m), 7.74 (1H, d, J = 1.8Hz), 7.93 (2H, d, J = 8.0 Hz), 7.98 (2H, d, J = 8.0 Hz), 8.20 (1H, d, J= 1.8 Hz), 10.04 (1H, s). MS (m/z): 242 (M + H)⁺. D-43

4-(4,5-dimethoxypyrimidin-2-yl)benzaldehyde ¹H-NMR (CDCl₃) δ: 4.00 (3H,s), 4.19 (3H, s), 7.97 (2H, d, J = 8.6 Hz), 8.17 (1H, s), 8.53 (2H, d, J= 8.6 Hz), 10.09 (1H, s). MS (m/z): 245 (M + H)⁺.

TABLE 1-5 D-44

3′-[(methanesulfonyl)methyl][1,1′-biphenyl]-4-carbaldehyde ¹H-NMR(CDCl₃) δ: 2.81 (3H, s), 4.31 (2H, s), 7.44-7.55 (2H, m), 7.66-7.68 (2H,m), 7.75 (2H, d, J = 8.3 Hz), 7.96 (2H, d, J = 8.3 Hz), 10.07 (1H, s).MS (m/z): 275 (M + H)⁺. D-45

4-[5-methoxy-6-(methylamino)pyridin-3-yl]benzaldehyde ¹H-NMR (CDCl₃) δ:3.09 (3H, d, J = 4.9 Hz), 3.93 (3H, s), 5.12 (1H, d, J = 4.9 Hz), 7.08(1H, d, J = 1.8 Hz), 7.66-7.71 (2H, m), 7.90-7.94 (2H, m), 8.09 (1H, d,J = 1.8 Hz), 10.02 (1H, s). MS (m/z): 243 (M + H)⁺. D-46

4-(5,6-dimethoxypyrazin-2-yl)benzaldehyde ¹H-NMR (CDCl₃) δ: 4.10 (3H,s), 4.15 (3H, s), 7.96 (2H, d, J = 8.5 Hz), 8.12 (2H, d, J = 7.9 Hz),8.20 (1H, s), 10.06 (1H, s). MS (m/z): 245 (M + H)⁺. D-47

4-[5-methoxy-6-(trifluoromethyl)pyridin-3yl]benzaldehyde ¹H-NMR (CDCl₃)δ: 4.11 (3H, s), 7.67-7.76 (2H, m), 7.96-8.04 (2H, m), 8.08-8.14 (1H,m), 8.55- 8.63 (1H, m), 10.08 (1H, s). MS (m/z): 282 (M + H)⁺.

TABLE 1-6 D-48

4-(5-ethoxy-6-methoxypyridin-3-yl)benzaldehyde ¹H-NMR (CDCl₃) δ: 1.53(3H, t, J = 7.0 Hz), 4.08 (3H, s), 4.19 (2H, q, J = 7.0 Hz), 7.28 (1H,d, J = 1.8 Hz), 7.68-7.72 (2H, m), 7.94-7.98 (2H, m), 8.01 (1H, d, J =1.8 HZ), 10.06 (1H, s). (m/z): 258 (M + H)⁺. D-49

N-(4′-formyl[1,1′-biphenyl]-3-yl)-N-methylmethanesulfonamide ¹H-NMR(CDCl₃) δ: 2.90 (3H, s), 3.40 (3H, s), 7.40 7.44 (1H, m), 7.49-7.54 (1H,m). 7.56-7.59 (1H, m), 7.66-7.68 (1H, m), 7.73-7.77 (2H, m), 7.95-7.99(2H, m). 10.07 (1H, s). MS (m/z): 290 (M + H)⁺. D-50

4-[6-(hydroxymethyl)-5′-methoxypyridin-3-yl]benzaldehyde ¹H-NMR (CDCl₃)δ: 3.95 (3H, s), 4.18 (1H, t, J = 4.9 Hz), 4.81 (2H, d, J = 4.9 Hz),7.33 (1H, s), 7.76 (2H, d, J = 8.6 Hz), 8.01 (2H, d, J = 8.6 Hz), 8.43(1H, s), 10.09 (1H, s). 13H D-51

4-[1-(methanesulfonyl)-1H-pyrrolol[2,3-c]pyridin-4-yl]benzaldehyde¹H-NMR (CDCl₃) δ: 3.28 (3H, s), 6.90 (1H, d, J = 3.7 Hz), 7.70 (1H, d, J= 3.7 Hz), 7.80 (2H, d, J = 8.6 Hz), 8.06 (2H, d, J = 8.6 Hz), 8.61 (1H,s), 9.30 (1H, s), 10.12 (1H, s).

TABLE 1-7 D-52

  CAS: 1446002-37-4

  CAS: 128376-64-74-[6-(methanesulfonyl)-5-methoxypyridin-3-yl]benzaldehyde ¹H-NMR(DMSO-D₆) δ: 3.35 (3H, s), 4.10 (3H, s), 8.08 (2H, d, J = 8.6 Hz), 8.13(2H, d, J = 8.6 Hz), 8.12 (1H, d, J = 1.8 Hz), 8.65 (1H, d, J = 1.8 Hz),10.11 (1H, s). MS (m/z): 292 (M + H)⁺. D-53

  Reference Example D-14

  CAS: 128376-64-74-[5-methoxy-6-(2-methoxyethoxy)pyridazin-3-yl]benzaldehyde ¹H-NMR(CDCl₃) δ: 3.47 (3H, s), 3.88 (2H, dd, J = 5.2, 4.0 Hz), 4.03 (3H, s),4.78-4.81 (2H, m), 7.19 (1H, s), 8.01 (2H, d, J = 8.6 Hz), 8.17 (2H, d,J = 8.6 Hz), 10.10 (1H, s). MS (m/z): 289 (M + H)⁺. D-54

  CAS: 1000342-04-0

  CAS: 128376-64-74-(4-chloro-1H-pyrrolo[3,2-c]pyridin-7-yl)benzaldehyde ¹H-NMR (DMSO-D₆)δ: 6.69 (1H, dd, J = 3.1, 1.8 Hz), 7.61 (1H, dd, J = 3.1, 2.5 Hz), 7.93(2H, d, J = 8.0 Hz), 8.09 (2H, d, J = 8.0 Hz), 8.11 (1H, s), 10.13 (1H,s), 12.08 (1H, br s). MS (m/z): 257 (M + H)⁺. D-55

  CAS: 1532-97-4

  CAS: 128376-64-7 4-(isoquinolin-4-yl)benzaldehyde ¹H-NMR (CDCl₃) δ:7.65-7.75 (4H, m), 7.87 (1H, 8.09 d, J = 8.0 Hz), 8.06 (2H, d, J = 8.0Hz), 8.09 (1H, d, J = 7.4 Hz), 8.51 (1H, s), 9.31 (1H, s), 10.14 (1H,s). MS (m/z): 234 (M + H)⁺.

TABLE 1-8 D-56

  CAS: 52605-98-8

  CAS: 128376-64-7 4-(5,6-dimethoxypyridin-3-yl)benzaldehyde ¹H-NMR(CDCl₃) δ: 3.97 (3H, s), 4.09 (3H, s), 7.28 (1H, d, J = 1.8 Hz), 7.71(2H, d, J = 8.6 Hz), 7.97 (2H, d, J = 8.6 Hz), 8.02 (1H, d, J = 1.8 Hz),10.06 (1H, s). MS (m/z): 244 (M + H)⁺. D-57

  Reference Example D-18

  CAS: 87199-17-54-[6-(azetidin-1-yl)-5-methoxypyridazin-3-yl]benzaldehyde ¹H-NMR (CDCl₃)δ: 2.43 (2H, quint, J = 7.4 Hz), 3.93 (3H, s), 4.35 (4H, t, J = 7.4 Hz),6.98 (1H, s), 7.97 (2H, d, J = 8.6 Hz), 8.16 (2H, d, J = 8.6 Hz), 10.07(1H, s). D-58

  Reference Example D-21

  CAS: 87199-17-54-[6-(difluoromethoxy)-5-methoxypyridazin-3-yl]benzaldehyde ¹H-NMR(CDCl₃) δ: 4.09 (3H, s), 7.33 (1H, s), 7.73 (1H, t, J = 71.7 Hz), 8.04(2H, d, J = 7.9 Hz), 8.17 (2H, d, J = 7.9 Hz), 10.12 (1H, s). MS (m/z) :281 (M + H)⁺. D-59

  Reference Example D-22 isomer A

  CAS: 87199-17-54-[6-(difluoromethoxy)-4-methoxypyridazin-3-yl]benzaldehyde ¹H-NMR(CDCl₃) δ: 4.00 (3H, s), 6.65 (1H, s), 7.75 (1H, t, J = 72.0 Hz),7.98-8.01 (2H, m), 8.05-8.08 (2H, m), 10.10 (1H, s). MS (m/z): 281 (M +H)⁺.

TABLE 1-9 D-60

  Reference Example D-22 (isomer B)

  CAS: 87199-17-54-[1-(difluoromethyl)-4-methoxy-6-oxo-1,6-dihydropyridazin-3-yl]benzaldehyde¹H-NMR (CDCl₃) δ: 3.93 (3H, s), 6.23 (1H, s), 7.74 (1H, t, J = 58.9 Hz),7.90 (2H, d, J = 8.2 Hz), 7.97 (2H, d, J = 8.2 Hz), 10.09 (1H, s) MS(m/z): 281 (M + H)⁺. D-61

  CAS: 1227502-46-6

  CAS: 87199-17-5 4-(4,5-dimethoxypyridin-2-yl)benzaldehyde ¹H-NMR(CDCl₃) δ: 4.02 (3H, s), 4.03 (3H, s), 7.30 (1H, s), 7.97 (2H, d, J =8.0 Hz), 8.10 (2H, d, J = 8.0 Hz), 8.26 (1H, s), 10.07 (1H, s). MS(m/z): 244 (M + H)⁺. D-62

  Reference Example D-15

  CAS: 87199-17-5 4-(6-{[1-(methanesulfonyl)piperidin-4-yl]oxy}-5-methoxypyridazin-3-yl)benzaldehyde MS (m/z): 392 (M + H)⁺. D-63

  CAS: 808770-39-0

  CAS: 87199-17-5 4-(6-amino-5-methoxypyridazin-3-yl)benzaldehyde ¹H-NMR(DMSO-D₆) δ: 3.99 (3H, s), 6.49 (2H, br s), 7.49 (1H, s), 8.00 (2H, d, J= 8.5 Hz), 8.29 (2H, d, J = 8.5 Hz), 10.06 (1H, s).

TABLE 1-10 D-64

  racemate Reference Example D-17

  CAS: 87199-17-5 racemate4-{5-methoxy-6-[(oxan-3-yl)oxy]pyridazin-3-yl}benzaldehyde ¹H-NMR(CDCl₃) δ: 1.70-1.72 (1H, m), 1.97-2.00 (2H, m), 2.23-2.25 (1H, m),3.71-3.73 (2H, m), 3.84-3.87 (1H, m), 4.02-4.03 (4H, m), 5.47-5.48 (1H,m), 7.19 (1H, s), 8.01 (2H, d, J = 8.3 Hz), 8.17 (2H, d, J = 8.3 Hz),10.10 (1H, s). MS (m/z): 315 (M + H)⁺. D-65

  CAS: 850991-69-4

  CAS: 15164-44-0 4-(5-methoxypyridin-3-yl)benzaldehyde ¹H-NMR (CDCl₃)δ: 3.95 (3H, s), 7.40-7.42 (1H, m), 7.73-7.78 (2H, m), 7.98-8.02 (2H,m), 8.37 (1H, d, J = 3.0 Hz), 8.50 (1H, d, J = 1.8 Hz), 10.09 (1H, s).D-66

  CAS: 832695-88-2

  CAS: 1122-91-4 4′-formyl-N-methyl[1,1′-biphenyl]-3-carboxamide ¹H-NMR(DMSO-D₆) δ: 3.06 (3H, s), 6.24 (1H, s), 7.54-7.56 (1H, m), 7.76-7.78(4H, m), 7.97- 7.98 (2H, m), 8.07-8.07 (1H, m), 10.07 (1H, s). MS (m/z):240 (M + H)⁺. D-67

  CAS: 871329-75-8

  CAS: 1122-91-4 4′-formyl-N-methyl[1,1′-biphenyl]-3-sulfonamide ¹H-NMR(CDCl₃) δ: 2.73 (3H, d, J = 5.5 Hz), 4.49-4.50 (1H, m), 7.65-7.67 (1H,m), 7.78 (2H, d, J = 8.3 Hz), 7.85-7.87 (1H, m), 7.91-7.93 (1H, m), 8.00(2H, d, J = 8.3 Hz), 8.14-8.14 (1H, m), 10.09 (1H, s). MS (m/z): 276(M + H)⁺.

TABLE 1-11 D-68

  CAS: 1451392-07-6

  CAS: 1122-91-4 4-(6-fluoro-5-methoxypyridin-3-yl)benzaldehyde ¹H-NMR(CDCl₃) δ: 4.00 (3H, s), 7.44-7.51 (1H, m), 7.69-7.75 (2H, m), 7.95-8.03(3H, m), 10.09 (1H, s). MS (m/z): 232 (M + H)⁺. D-69

  CAS: 1451392-07-6

  CAS: 133059-43-53-fluoro-4-(6-fluoro-5-methoxypyridin-3-yl)benzaldehyde ¹H-NMR (CDCl₃)δ: 3.98 (3H, s), 7.49-7.53 (1H, m), 7.61-7.66 (1H, m), 7.70- 7.73 (1H,m), 7.78-7.80 (1H, m), 7.93-7.95 (1H, m), 10.05 (1H, d, J = 1.8 Hz).D-70

  CAS: 609807-25-2

  CAS: 15164-44-0 3′-fluoro-5′-methoxy[1,1′-biphenyl]-4-carbaldehyde¹H-NMR (CDCl₃) δ: 3.87 (3H, s), 6.68 (1H, dt, J = 10.3, 2.1 Hz),6.92-6.95 (2H, m), 7.72 (2H, d, J = 8.5 Hz), 7.96 (2H, d, J = 8.5 Hz),10.07 (1H, s). D-71

  CAS: 850991-69-4

  CAS: 149806-06-4 5′-methoxy[2,3′-bipyridine]-5-carbaldehyde ¹H-NMR(CDCl₃) δ: 3.97 (3H, s), 7.96 (1H, d, J = 8.0 Hz), 7.99 (1H, dd, J =3.1, 1.8 Hz), 8.27 (1H, dd, J = 8.0, 1.8 Hz), 8.43 (1H, d, J = 3.1 Hz),8.85 (1H, d, J = 1.8 Hz), 9.16 (1H, d, J = 1.8 Hz), 10.17 (1H, s). MS(m/z): 215 (M + H)⁺.

TABLE 1-12 D-72

   

    CAS: 1083168-92-6  

  CAS: 24078-12-4 4-(5,6-dimethoxypyridin-3-yl)-2-methylbenzaldehyde¹H-NMR (CDCl₃) δ: 2.75 (3H, s), 3.97 (3H, s), 4.08 (3H, s), 7.27 (1H, d,J = 2.5 Hz), 7.43 (1H, s), 7.53 (1H, d, J = 8.0 Hz), 7.88 (1H, d, J =8.0 Hz), 8.01 (1H, d, J = 2.5 Hz), 10.30 (1H, s). MS (m/z): 258 (M +H)⁺. D-73

  CAS: 1083168-92-6

  CAS: 43192-33-2 4-(5,6-dimethoxypyridin-3-yl)-2-methoxybenzaldehyde¹H-NMR (CDCl₃) δ: 3.97 (3H, s), 4.01 (3H, s), 4.08 (3H, s), 7.11 (1H, d,J = 1.8 Hz), 7.19 (1H, dd, J = 8.0, 1.8 Hz), 7.25 (1H, d, J = 1.8 Hz),7.91 (1H, d, J = 8.0 Hz), 8.00 (1H, d, J = 1.8 Hz), 10.48 (1H, s). MS(m/z): 274 (M + H)⁺. D-74

  CAS: 1083168-92-6

  CAS: 57848-46-1 4-(5,6-dimethoxypyridin-3-yl)-2-fluorobenzaldehyde¹H-NMR (CDCl₃) δ: 3.97 (3H, s), 4.09 (3H, s), 7.24 (1H, d, J = 1.8 Hz),7.35 (1H, d, J = 11.7 Hz), 7.46 (1H, d, J = 8.6 Hz), 7.95 (1H, t, J =7.7 Hz), 8.01 (1H, d, J = 1.2 Hz), 10.39 (1H, s). MS (m/z): 262 (M +H)⁺.

Reference Example D-754-[1-(methanesulfonyl)-1H-pyrazol-4-yl]benzaldehyde Step 14-(1H-pyrazol-4-yl)benzaldehyde hydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in Reference ExampleD-27.

¹H-NMR (DMSO-D₆) δ: 7.86-7.90 (4H, m), 8.29 (2H, s), 9.96 (1H, s), 11.97(2H, s).

MS (m/z): 173 (M+H)⁺.

Step 2 4-[1-(methanesulfonyl)-1H-pyrazol-4-yl]benzaldehyde

The title compound was obtained in the same manner as in ReferenceExample D-3, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 3.42 (3H, s), 7.70 (2H, d, J=8.3 Hz), 7.95 (2H, d,J=8.3 Hz), 8.17 (1H, s), 8.38 (1H, s), 10.03 (1H, s).

Reference Example D-76 4-(5-methoxy-6-phenoxypyridazin-3-yl)benzaldehyde

The title compound was obtained in the same manner as in Step 1 ofReference Example D-16, using the compound obtained in Reference ExampleD-28 and phenol.

¹H-NMR (CDCl₃) δ: 4.12 (3H, s), 7.27-7.28 (2H, m), 7.33 (1H, s),7.41-7.44 (2H, m), 8.00-8.01 (2H, m), 8.04-8.05 (1H, m), 8.18-8.20 (2H,m), 10.09 (1H, s).

MS (m/z): 307 (M+H)⁺.

Reference Example D-774′-formyl-N,N-dimethyl[1,1′-biphenyl]-3-sulfonamide

The title compound was obtained in the same manner as in Step 13 ofReference Example A-1, using the compound obtained in Reference ExampleD-67

1H-NMR (CDCl₃) δ: 2.77 (6H, s), 7.71-7.84 (5H, m), 8.00-8.03 (3H, m),10.09 (1H, s).

MS (m/z): 290 (M+H)⁺.

Reference Example D-784-(5,6-dimethoxypyridazin-3-yl)cyclohexane-1-carbaldehyde (cis-transmixture) Step 14-(5,6-dimethoxypyridazin-3-yl)cyclohex-3-ene-1-carbaldehyde (racemate)

The compound (0.400 g) obtained in Reference Example D-35 was dissolvedin dichloromethane (11 mL), and diisobutylaluminium hydride (1.22 mol/L,dichloromethane solution, 2.24 mL) was added dropwise thereto at −78° C.The reaction solution was stirred at the same temperature for 2 hr.Methanol (0.400 mL) was added to the reaction solution, and thensaturated aqueous potassium sodium tartrate solution was added thereto,and the mixture was allowed to warmed to room temperature. The mixturewas extracted with ethyl acetate, the organic layer was dried overanhydrous sodium sulfate, and the solvent was evaporated under reducedpressure. The residue was purified by silica gel chromatography(n-hexane/ethyl acetate) to give the title compound (0.178 g) as asolid.

¹H-NMR (CDCl₃) δ: 1.79-1.89 (1H, m), 2.19-2.27 (1H, m), 2.43-2.77 (4H,m), 2.81-2.90 (1H, m), 3.95 (3H, s), 4.18 (3H, s), 6.44-6.49 (1H, m),6.90 (1H, s), 9.78 (1H, s).

MS (m/z): 249 (M+H)⁺.

Step 2 4-(5,6-dimethoxypyridazin-3-yl)cyclohexane-1-carbaldehyde(cis-trans mixture)

The compound (0.175 g) obtained in the above Step 1 was dissolved inethyl acetate (7 mL), 10% palladium on carbon wet (0.300 g) was addedthereto, and the mixture was stirred under hydrogen atmosphere at roomtemperature for 5 hr. The reaction solution was filtered, and thefiltrate was concentrated under reduced pressure to give the titlecompound (0.179 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.39-2.94 (10H, m), 3.91-3.94 (3H, m), 4.14-4.16 (3H,m), 6.54-6.60 (1H, m), 9.69-9.78 (1H, m).

Reference Example D-79N-[6-(4-formylphenyl)-4-methoxypyridazin-3-yl]-N-methylprop-2-enamideStep 1 4-[5-methoxy-6-(methylamino)pyridazin-3-yl]benzaldehyde

A mixture of the compound (100 mg) obtained in Reference Example D-63,methyl iodide (0.032 mL), potassium carbonate (121 mg) and DMF (2.2 mL)was stirred at 70° C. for 1.5 hr. Water was added to the reactionsolution, and the precipitated solid was collected by filtration to givethe title compound (62.3 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.73 (3H, s), 4.00 (3H, s), 7.03 (1H, s), 7.99 (2H,d, J=8.3 Hz), 8.13 (2H, d, J=8.3 Hz), 10.05 (1H, s).

MS (m/z): 244 (M+H)⁺.

Step 2N-[6-(4-formylphenyl)-4-methoxypyridazin-3-yl]-N-methylprop-2-enamide

To a mixture of the compound (62.3 mg) obtained in the above Step 1,DIPEA (0.134 mL) and dichloromethane (2.56 mL) was added acryloylchloride (0.0228 mL) under ice-cooling, and the mixture was stirred atroom temperature for 1 hr. Water was added to the reaction solution, andthe mixture was extracted with dichloromethane. The organic layer waswashed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (ethyl acetate/methanol) to give the titlecompound (43.4 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 3.86 (3H, s), 3.94 (3H, s), 5.63 (1H, dd, J=10.0,2.1 Hz), 6.00 (1H, dd, J=17.3, 2.1 Hz), 6.26 (1H, dd, J=17.3, 10.0 Hz),7.46 (1H, s), 8.04 (2H, d, J=8.2 Hz), 8.22 (2H, d, J=8.2 Hz), 10.09 (1H,s).

MS (m/z): 298 (M+H)⁺.

Reference Example D-80 4-[(pyridin-3-yl)amino]benzaldehyde

A mixture of 4-bromobenzaldehyde (150 mg), pyridin-3-amine (91.6 mg),tris(dibenzylideneacetone)dipalladium(0) (37.1 mg),4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (51.6 mg), cesiumcarbonate (370 mg) and 1,4-dioxane (1.62 mL) was stirred overnight undernitrogen atmosphere at 100° C. The reaction solution was cooled, andfiltered through Celite, and the filtrate was concentrated under reducedpressure. The residue was purified by silica gel column chromatography(ethyl acetate/methanol) to give the title compound (146 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 7.15 (2H, d, J=8.6 Hz), 7.36-7.38 (1H, m), 7.65-7.67(1H, m), 7.77 (2H, d, J=8.6 Hz), 8.22-8.25 (1H, m), 8.47-8.48 (1H, m),9.13 (1H, s), 9.75 (1H, s).

MS (m/z): 199 (M+H)⁺.

Reference Example D-81 3-[(pyridin-3-yl)amino]benzaldehyde

The title compound was obtained in the same manner as in ReferenceExample D-80 except that 3-bromobenzaldehyde was used instead of4-bromobenzaldehyde.

¹H-NMR (CDCl₃) δ: 6.04 (1H, s), 7.22-7.33 (2H, m), 7.44-7.48 (3H, m),7.55-7.56 (1H, m), 8.25 (1H, dd, J=4.6, 1.5 Hz), 8.44 (1H, d, J=2.4 Hz),9.96 (1H, s).

MS (m/z): 199 (M+H)⁺.

Reference Example D-82 4-(2,3-dihydro-1H-indol-1-yl)benzaldehyde Step 11-[4-(1,3-dioxolan-2-yl)phenyl]-2,3-dihydro-1H-indole

A mixture of indoline (0.280 mL), 2-(4-bromophenyl)-1,3-dioxolane (CAS:10602-01-4) (1.15 g), TEA (1.15 mL),(±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (54.9 mg),tris(dibenzylideneacetone)dipalladium(0) (69.2 mg), cesium carbonate(2.46 g) and toluene (20 mL) was stirred under nitrogen atmosphere at110° C. for 5 hr. Saturated aqueous sodium hydrogencarbonate solutionwas added to the reaction solution, and the mixture was extracted withethyl acetate. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure,and the residue was subjected to silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (342 mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.13 (2H, t, J=8.3 Hz), 3.96 (2H, t, J=8.3 Hz),4.02-4.06 (2H, m), 4.14-4.18 (2H, m), 5.78 (1H, s), 6.76 (1H, td, J=7.4,1.2 Hz), 7.07 (1H, t, J=7.4 Hz), 7.14-7.18 (2H, m), 7.22 (2H, d, J=8.6Hz), 7.45 (2H, d, J=8.6 Hz).

Step 2 4-(2,3-dihydro-1H-indol-1-yl)benzaldehyde

To a solution of the compound (155 mg) obtained in the above Step 1 inTHF (2.3 mL) was added 1N hydrochloric acid (2.32 mL) at roomtemperature, and the mixture was stirred at the same temperature for 16hr. Saturated aqueous sodium hydrogencarbonate solution was added to thereaction solution, and the mixture was extracted with chloroform. Theorganic layer was washed with saturated brine, dried over anhydroussodium sulfate, and concentrated under reduced pressure, and the residuewas subjected to silica gel column chromatography (n-hexane/ethylacetate) to give the title compound (115 mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.19 (2H, t, J=8.3 Hz), 4.06 (2H, t, J=8.3 Hz), 6.89(1H, t, J=8.0 Hz), 7.16 (1H, t, J=8.0 Hz), 7.24 (1H, d, J=8.0 Hz), 7.28(2H, d, J=9.2 Hz), 7.35 (1H, d, J=8.0 Hz), 7.84 (2H, d, J=9.2 Hz), 9.84(1H, s).

MS (m/z): 224 (M+H)⁺.

Reference Example D-83 N-[3-(4-formylanilino)phenyl]prop-2-enamide Step1 N-[4-(1,3-dioxolan-2-yl)phenyl]-3-nitroaniline

A mixture of 3-nitroaniline (CAS: 99-09-2) (4.13 g),2-(4-bromophenyl)-1,3-dioxolane (5.71 g), sodium tert-butoxide (CAS:865-48-5) (4.31 g), tris(dibenzylideneacetone)dipalladium(0) (1.14 g),tri-tert-butylphosphine (10% n-hexane solution) (CAS: 13716-12-6) (7.42mL) and toluene (50 mL) was stirred at room temperature for 1 hr, andthen at 80° C. for 1 hr. Ethyl acetate and water were added to thereaction solution, and the mixture was filtered through Celite. Thefiltrate was subjected to liquid separation, and the aqueous layer wasextracted with ethyl acetate. The organic layers were combined, washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby amino silica gel column chromatography (n-hexane/ethyl acetate). Thesolvent was evaporated under reduced pressure, a mixed solvent ofn-hexane/diethyl ether=1/1 was added to the residue, and the solid wascollected by filtration to give the title compound (4.08 g) as a solid.

¹H-NMR (CDCl₃) δ: 4.03-4.10 (2H, m), 4.11-4.18 (2H, m), 5.78 (1H, s),6.02 (1H, br s), 7.12-7.15 (2H, m), 7.30 (1H, dd, J=7.9, 2.4 Hz), 7.38(1H, t, J=7.9 Hz), 7.44-7.48 (2H, m), 7.70-7.73 (1H, m), 7.87 (1H, t,J=2.4 Hz).

Step 2 N¹-[4-(1,3-dioxolan-2-yl)phenyl]benzene-1,3-diamine

To the compound (1.50 g) obtained in the above Step 1 and 10% palladiumon carbon wet (0.300 g) was added ethanol (20 mL), and the mixture wasstirred under hydrogen atmosphere at room temperature for 2 hr. Thepalladium was removed by filtration, and the filtrate was concentratedunder reduced pressure to give the title compound (1.29 g) as a solid.This was used in the next reaction without further purification.

¹H-NMR (CDCl₃) δ: 3.62 (2H, br s), 3.99-4.07 (2H, m), 4.10-4.19 (2H, m),5.69 (1H, br s), 5.74 (1H, s), 6.29 (1H, dd, J=7.9, 1.8 Hz), 6.44 (1H,t, J=1.8 Hz), 6.47 (1H, dd, J=7.9, 1.8 Hz), 7.03-7.07 (3H, m), 7.34-7.38(2H, m).

Step 3 N-{3-[4-(1,3-dioxolan-2-yl)anilino]phenyl}prop-2-enamide

The compound (590 mg) obtained in the above Step 2 was dissolved indichloromethane (15 mL), and DIPEA (1.2 mL) and acryloyl chloride (CAS:814-68-6) (0.205 mL) were added thereto, and the mixture was stirred atroom temperature for 1 hr. Dichloromethane and water was added thereto,and the mixture was subjected to liquid separation. The organic layerwas dried over anhydrous sodium sulfate, and concentrated under reducedpressure. The obtained residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (709mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.99-4.08 (2H, m), 4.10-4.19 (2H, m), 5.73-5.77 (2H,m), 5.83 (1H, br s), 6.22 (1H, dd, J=17.0, 10.3 Hz), 6.42 (1H, dd,J=17.0, 1.2 Hz), 6.84 (1H, dd, J=8.5, 1.8 Hz), 7.04-7.09 (3H, m), 7.20(1H, t, J=8.5 Hz), 7.35-7.42 (4H, m).

Step 4 N-[3-(4-formylanilino)phenyl]prop-2-enamide

The title compound was obtained in the same manner as in Step 2 ofReference Example D-82, using the compound obtained in the above Step 3.

1H-NMR (DMSO-D₆) δ: 5.77 (1H, dd, J=10.3, 1.5 Hz), 6.26 (1H, dd, J=17.0,1.5 Hz), 6.44 (1H, dd, J=17.0, 10.3 Hz), 6.91 (1H, d, J=7.9 Hz), 7.15(2H, d, J=8.5 Hz), 7.24-7.31 (2H, m), 7.72-7.76 (3H, m), 9.05 (1H, s),9.73 (1H, s), 10.17 (1H, s).

Reference Example D-844-formyl-N-[(1-methyl-1H-pyrazol-4-yl)methyl]benzamide

A mixture of 4-formylbenzoic acid (CAS: 619-66-9) (150 mg),(1-methyl-1H-pyrazol-4-yl)methanamine (122 mg), HOBt (162 mg),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (WSCD) (230mg) and DMF (8.0 mL) was stirred at room temperature for 15 hr. Waterwas added to the reaction solution, and the mixture was extracted withethyl acetate. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure,and the residue was subjected to silica gel column chromatography(chloroform/methanol) to give the title compound (200 mg) as a solid.

¹H-NMR (CDCl₃) δ: 3.83 (3H, s), 4.48 (2H, d, J=6.1 Hz), 6.12 (1H, br s),7.38-7.47 (2H, m), 7.69-7.82 (1H, m), 7.88-7.93 (3H, m), 10.03 (1H, s).

MS (m/z): 244 (M+H)⁺.

Reference Example D-85N-{3-[(ethenesulfonyl)amino]propyl}-4-formylbenzamide Step 1 tert-butyl[3-(4-formylbenzamido)propyl]carbamate

The title compound was obtained in the same manner as in ReferenceExample D-84 except that tert-butyl N-(3-aminopropyl)carbamate (CAS:75178-96-0) was used instead of (1-methyl-1H-pyrazol-4-yl)methanamine.

¹H-NMR (CDCl₃) δ: 1.46 (9H, s), 1.71-1.77 (2H, m), 3.25-3.29 (2H, m),3.51-3.55 (2H, m), 5.01 (1H, br s), 7.75 (1H, br s), 7.96 (2H, d, J=7.9Hz), 8.04 (2H, d, J=7.9 Hz), 10.09 (1H, s).

MS (m/z): 207 (M−Boc+H)⁺.

Step 2 N-{3-[(ethenesulfonyl)amino]propyl}-4-formylbenzamide

The compound (449 mg) obtained in the above Step 1 was dissolved indichloromethane (3 mL), and hydrogen chloride (4 mol/L, 1,4-dioxanesolution, 3 mL) was added thereto. The mixture was stirred at roomtemperature for 2 hr, and the solvent was evaporated under reducedpressure. Dichloromethane (11 mL), DIPEA (1.92 mL) and2-chloroethanesulfonyl chloride (CAS: 1622-32-8) (0.174 mL) were addedto the residue, and the mixture was stirred at room temperature for 3hr. The solvent was evaporated under reduced pressure. The residue waspurified by silica gel column chromatography (ethyl acetate) to give thetitle compound (110 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.85 (2H, quint. J=6.1 Hz), 3.12 (2H, q, J=6.1 Hz),3.62 (2H, q, J=6.1 Hz), 5.57 (1H, t, J=6.1 Hz), 5.94 (1H, d, J=9.7 Hz),6.22 (1H, d, J=17.0 Hz), 6.55 (1H, dd, J=17.0, 9.7 Hz), 7.16 (1H, t,J=6.1 Hz), 7.91-7.96 (4H, m), 10.06 (1H, s).

MS (m/z): 297 (M+H)+.

Reference Example D-86 4-[(2-oxopyrrolidin-1-yl)methyl]benzaldehyde Step1 methyl 4-[(2-oxopyrrolidin-1-yl)methyl]benzoate

To a suspension of sodium hydride (purity 55%, 238 mg) and DMF (15 mL)was added 2-pyrrolidinone (CAS: 616-45-5) (557 mg) under ice-cooling,and after 30 min, a solution of methyl 4-(bromomethyl)benzoate (CAS:2417-72-3) (1.00 g) in DMF (15 mL) was added dropwise thereto at thesame temperature. The mixture was stirred at room temperature for 16 hr,water was added to the reaction solution, and the mixture was extractedwith ethyl acetate. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure, and the residue was subjected to silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (576mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.99-2.06 (2H, m), 2.47 (2H, t, J=7.9 Hz), 3.27 (2H,t, J=7.3 Hz), 3.92 (3H, s), 4.51 (2H, s), 7.31 (2H, d, J=8.5 Hz), 8.01(2H, d, J=8.5 Hz).

MS (m/z): 234 (M+H)⁺.

Step 2 1-{[4-(hydroxymethyl)phenyl]methyl}pyrrolidin-2-one

To a solution of the compound (576 mg) obtained in the above Step 1 inTHF (20 mL) was added lithium borohydride (CAS: 16949-15-8) (108 mg)little by little under ice-cooling, and methanol (0.2 mL) was addedthereto at the same temperature. The mixture was stirred at roomtemperature for 18 hr, and then at 50° C. for 4 hr. Lithium borohydride(108 mg) was again added little by little under ice-cooling to thereaction solution, and the mixture was stirred at 60° C. for 9 hr. Thereaction solution was weakly acidified with 1N hydrochloric acid byaddition little by little under ice-cooling, and extracted withchloroform. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure,and the residue was subjected to silica gel column chromatography(chloroform/methanol) to give the title compound (338 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.95-2.03 (2H, m), 2.44 (2H, t, J=7.9 Hz), 3.25 (2H,t, J=7.3 Hz), 4.44 (2H, s), 4.69 (2H, s), 7.23 (2H, d, J=7.9 Hz), 7.34(2H, d, J=7.9 Hz).

MS (m/z): 206 (M+H)⁺.

Step 3 4-[(2-oxopyrrolidin-1-yl)methyl]benzaldehyde

To a solution of the compound (336 mg) obtained in the above Step 2 indichloromethane (15 mL) was added Dess-Martin periodinane (CAS:87413-09-0) (764 mg) under ice-cooling. The mixture was stirred at roomtemperature for 1.5 hr, water was added to the reaction solution, andthe mixture was extracted with ethyl acetate. The organic layer waswashed with saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure, and the residue was subjected tosilica gel column chromatography (chloroform/methanol) to give the titlecompound (343 mg) as an oil.

¹H-NMR (CDCl₃) δ: 2.01-2.08 (2H, m), 2.48 (2H, t, J=8.6 Hz), 3.30 (2H,t, J=7.4 Hz), 4.54 (2H, s), 7.41 (2H, d, J=8.0 Hz), 7.86 (2H, d, J=8.0Hz), 10.01 (1H, s).

MS (m/z): 204 (M+H)⁺.

Reference Example D-87 4-(5-oxopyrrolidin-2-yl)benzaldehyde(racemate)Step 1 methyl 4-(5-oxopyrrolidin-2-yl)benzoate (racemate)

A mixture of 5-(4-bromophenyl)pyrrolidin-2-one (CAS: 207989-90-0) (1.00g), TEA (1.15 mL), methanol (15 mL),[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloridedichloromethane adduct (170 mg) and DMF (30 mL) was stirred under carbonmonoxide atmosphere at 90° C. for 2 hr. Aqueous saturated ammoniumchloride solution was added to the reaction solution, and the mixturewas extracted with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure, and the residue was subjected to silica gelcolumn chromatography (chloroform/methanol) to give the title compound(342 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.91-2.02 (1H, m), 2.41-2.50 (2H, m), 2.56-2.67 (1H,m), 3.93 (3H, s), 4.82 (1H, t, J=7.4 Hz), 5.96-6.18 (1H, m), 7.38 (2H,d, J=8.0 Hz), 8.05 (2H, d, J=8.0 Hz).

MS (m/z): 220 (M+H)⁺.

Step 2 5-[4-(hydroxymethyl)phenyl]pyrrolidin-2-one (racemate)

The title compound was obtained in the same manner as in Step 2 ofReference Example D-86, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.67-1.77 (1H, m), 2.22 (2H, t, J=8.0 Hz), 2.39-2.47(1H, m), 4.47 (2H, d, J=5.5 Hz), 4.64 (1H, t, J=7.1 Hz), 5.15 (1H, t,J=5.8 Hz), 7.24 (2H, d, J=8.0 Hz), 7.29 (2H, d, J=8.0 Hz), 8.07 (1H, brs).

MS (m/z): 192 (M+H)⁺.

Step 3 4-(5-oxopyrrolidin-2-yl)benzaldehyde(racemate)

The compound (110 mg) obtained in the above Step 2 was dissolved inchloroform (5 mL), manganese dioxide (498 mg) was added thereto, and themixture was stirred at room temperature for 1.5 hr. Ethyl acetate andwater were added thereto, the insoluble substance was removed byfiltration, and the filtrate was subjected to liquid separation. Theaqueous layer was extracted with ethyl acetate, and the organic layerswere combined, washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The obtained residuewas purified by silica gel column chromatography (chloroform/methanol)to give the title compound (56 mg) as an oil.

¹H-NMR (CDCl₃) δ: 1.93-2.03 (1H, m), 2.44-2.52 (2H, m), 2.60-2.71 (1H,m), 4.85 (1H, t, J=7.4 Hz), 6.04 (1H, br s), 7.48 (2H, d, J=8.0 Hz),7.91 (2H, d, J=8.0 Hz), 10.02 (1H, s).

MS (m/z): 190 (M+H)⁺.

Reference Example D-88 4-(cyclohexylamino)benzaldehyde Step 1 methyl4-(cyclohexylamino)benzoate

A mixture of methyl 4-aminobenzoate (CAS: 619-45-4) (1.13 g), magnesiumsulfate (1.81 g), cyclohexanone (3.01 g), acetic acid (7.58 mL), sodiumtriacetoxyborohydride (14.3 g), methanol (30 mL) and dichloromethane (10mL) was stirred at room temperature for 36 hr, and then at 60° C. for 9hr. Saturated aqueous sodium hydrogencarbonate solution was added to thereaction mixture, and the mixture was extracted with chloroform. Theorganic layer was washed with saturated brine, dried over anhydroussodium sulfate, filtered, and concentrated under reduced pressure. Theobtained residue was subjected to silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (1.16 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.12-1.28 (3H, m), 1.32-1.44 (2H, m), 1.61-1.70 (1H,m), 1.77 (2H, dt, J=13.5, 3.7 Hz), 2.05 (2H, dd, J=12.9, 3.1 Hz), 3.31(1H, br s), 3.84 (3H, s), 4.03 (1H, br s), 6.50-6.54 (2H, m), 7.81-7.85(2H, m).

MS (m/z): 234 (M+H)⁺.

Step 2 [4-(cyclohexylamino)phenyl]methanol

The title compound was obtained in the same manner as in Step 2 ofReference Example D-86, using the compound obtained in the above Step 1.

Step 3 4-(cyclohexylamino)benzaldehyde

The title compound was obtained in the same manner as in Step 3 ofReference Example D-87, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.15-1.29 (3H, m), 1.34-1.45 (2H, m), 1.68 (1H, dt,J=12.9, 3.7 Hz), 1.79 (2H, dt, J=13.5, 3.7 Hz), 2.03-2.10 (2H, m),3.32-3.41 (1H, m), 4.25 (1H, d, J=8.0 Hz), 6.58 (2H, d, J=8.6 Hz), 7.67(2H, d, J=8.6 Hz), 9.70 (1H, s).

MS (m/z): 204 (M+H)⁺.

Reference Example D-89 tert-butyl benzyl (4-formylphenyl)carbamate Step1 methyl 4-[benzyl (tert-butoxycarbonyl)amino]benzoate

To a suspension of sodium hydride (purity 55%, 131 mg) and THF (10 mL)was added a solution of methyl 4-[(tert-butoxycarbonyl)amino]benzoate(503 mg) synthesized according to the method described in a literature(J. Med. Chem. 2016, 59(18), 8233-8262) in THF (5 mL) under ice-cooling.The mixture was stirred at room temperature for 30 min, and benzylbromide (CAS: 100-39-0) (0.310 mL) was added dropwise thereto underice-cooling. The mixture was stirred at room temperature for 15 hr,water was added to the reaction solution, and the mixture was extractedwith ethyl acetate. The organic layer was washed with saturated brine,dried over anhydrous sodium sulfate, and concentrated under reducedpressure, and the residue was subjected to silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (408mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.42 (9H, s), 3.89 (3H, s), 4.88 (2H, s), 7.19-7.32(7H, m), 7.94 (2H, d, J=8.6 Hz).

MS (m/z): 242 (M-Boc+H)⁺.

Step 2 tert-butyl benzyl [4-(hydroxymethyl)phenyl]carbamate

To a solution of the compound (405 mg) obtained in the above Step 1 inTHF (10 mL) was added lithium borohydride (CAS: 16949-15-8) (78.9 mg)little by little under ice-cooling. The mixture was stirred at roomtemperature for 1 hr, and the mixture was stirred at 50° C. for 4 hr.The reaction solution was weakly acidified with 1N hydrochloric acid byaddition little by little under ice-cooling, and extracted withchloroform. The organic layer was washed with saturated brine, and driedover anhydrous sodium sulfate to give a crude product (369 mg) of thetitle compound as an oil.

MS (m/z): 258 (M+H−tBu)⁺.

Step 3 tert-butyl benzyl (4-formylphenyl)carbamate

The title compound was obtained in the same manner as in Step 3 ofReference Example D-87, using the compound obtained in the above Step 2.

¹H-NMR (CDCl₃) δ: 1.43 (9H, s), 4.92 (2H, s), 7.22 (2H, d, J=6.7 Hz),7.27-7.35 (3H, m), 7.39 (2H, d, J=8.6 Hz), 7.79 (2H, d, J=8.6 Hz), 9.93(1H, s).

MS (m/z): 212 (M-Boc+H)⁺.

Reference Example D-901-(5,6-dimethoxypyridazin-3-yl)piperidine-4-carbaldehyde Step 1[1-(5,6-dimethoxypyridazin-3-yl)piperidin-4-yl]methanol

A mixture of 6-chloro-3,4-dimethoxypyridazine (364 mg), 4-piperidinemethanol (CAS: 586-95-8, 200 mg),(+/−)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (216 mg), sodiumtert-butoxide (200 mg), toluene (4 mL) andtris(dibenzylideneacetone)dipalladium(0) (159 mg) was stirred undernitrogen atmosphere at 100° C. for 4 hr. Water was added to the reactionsolution, and the mixture was extracted with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (ethyl acetate/methanol) togive the title compound (208 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.35-1.38 (2H, m), 1.75-1.78 (1H, m), 1.83-1.86 (2H,m), 2.89-2.93 (2H, m), 3.54-3.55 (2H, m), 3.90 (3H, s), 4.08 (3H, s),4.18-4.19 (1H, m), 4.20-4.23 (1H, m), 5.30 (1H, s), 6.35 (1H, s).

MS (m/z): 254 (M+H)⁺.

Step 2 1-(5,6-dimethoxypyridazin-3-yl)piperidine-4-carbaldehyde

The title compound was obtained in the same manner as in Step 3 ofReference Example D-86, using the compound obtained in the above Step 1.

1H-NMR (CDCl₃) δ: 1.70-1.79 (2H, m), 2.02-2.04 (2H, m), 2.50-2.53 (1H,m), 3.08-3.15 (2H, m), 3.90 (3H, s), 4.06-4.07 (5H, m), 6.35 (1H, s),9.70 (1H, s).

MS (m/z): 252 (M+H)⁺.

Reference Example D-91 4-(1H-1,2,4-triazol-5-yl)benzaldehyde Step 1[4-(1H-1,2,4-triazol-5-yl)phenyl]methanol

The title compound was obtained in the same manner as in Step 2 ofReference Example D-86, using 4-(1H-1,2,4-triazol-5-yl)benzoic acid(CAS: 876715-40-1). The obtained compound was directly used in the nextstep without purification.

MS (m/z): 176 (M+H)⁺.

Step 2 4-(1H-1,2,4-triazol-5-yl)benzaldehyde

A mixture of the compound (155 mg) obtained in the above Step 1,pyridinium chlorochromate (381 mg), DMSO (3 mL) and dichloromethane (8mL) was stirred overnight at room temperature. Saturated aqueous sodiumhydrogencarbonate solution was added to the reaction solution, and themixture was extracted with ethyl acetate. The organic layer was washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (ethyl acetate) to give the title compound(118 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 8.03 (2H, d, J=8.0 Hz), 8.25 (2H, d, J=8.0 Hz), 8.71(1H, br s), 10.06 (1H, s), 14.37 (1H, br s).

MS (m/z): 174 (M+H)⁺.

Reference Example D-924-(3-hydroxypiperidine-1-sulfonyl)benzaldehyde(racemate)

To a solution of 3-hydroxypiperidine (51.4 mg) in dichloromethane (1 mL)was added a solution of 4-formylbenzene-1-sulfonyl chloride (80 mg) indichloromethane (1 mL), and the mixture was stirred at room temperaturefor 1 hr. Water was added to the reaction solution, and the mixture wasextracted with dichloromethane. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (n-hexane/ethyl acetate) to give the title compound (102mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.40-1.44 (1H, m), 1.63-1.67 (1H, m), 1.76-1.95 (3H,m), 2.73-2.87 (2H, m), 3.18-3.23 (1H, m), 3.40 (1H, m), 3.87-3.89 (1H,m), 7.94 (2H, m), 8.05-8.07 (2H, m), 10.12 (1H, s).

MS (m/z): 270 (M+H)⁺.

Reference Example D-93 4-(azepane-1-sulfonyl)benzaldehyde

The title compound was obtained in the same manner as in ReferenceExample D-92 except that azepane was used instead of3-hydroxypiperidine.

¹H-NMR (CDCl₃) δ: 1.58-1.61 (4H, m), 1.72-1.75 (4H, m), 3.30-3.31 (4H,m), 7.96-8.02 (4H, m), 10.10 (1H, s).

MS (m/z): 268 (M+H)⁺.

Reference Example D-946-(4-formylphenyl)-4-methoxypyridazine-3-carbonitrile

A mixture of the compound (0.200 g) obtained in Reference Example D-28,zinc cyanide (0.0965 g), tetrakis(triphenylphosphine)palladium(0)(0.0930 g) and N,N-dimethylacetamide (3.00 mL) was stirred undernitrogen atmosphere at 100° C. for 1 hr. The reaction solution wasdiluted with ethyl acetate, washed successively with water and saturatedbrine, dried over anhydrous magnesium sulfate, and concentrated underreduced pressure. The obtained residue was subjected to silica gelcolumn chromatography (dichloromethane/ethyl acetate). The fraction wasconcentrated under reduced pressure, and the obtained solid wassubjected to slurry washing with dichloromethane, and dried to give thetitle compound (0.0826 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 4.20 (3H, s), 8.14 (2H, d, J=8.5 Hz), 8.21 (1H, s),8.51 (2H, d, J=8.5 Hz), 10.15 (1H, s).

MS (m/z): 240 (M+H)⁺.

Reference Example D-95 {4-[(5-methoxypyridin-3-yl)amino]phenyl}methanolStep 1N-[4-({[tert-butyl(dimethyl)silyl]oxy}methyl)phenyl]-5-methoxypyridin-3-amine

To 4-({[tert-butyl(dimethyl)silyl]oxy}methyl)aniline (0.404 g)synthesized according to the method described in a literature(Tetrahedron 2015, 71(49), 9240-9244), 3-bromo-5-methoxypyridine (CAS:50720-12-2) (0.320 g), tris(dibenzylideneacetone)dipalladium(0) (0.156g), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (CAS: 161265-03-8)(0.197 g) and cesium carbonate (1.67 g) was added 1,4-dioxane (2.43 mL),and the mixture was stirred under nitrogen atmosphere at 70° C. for 2hr. The reaction mixture was concentrated under reduced pressure, andthe residue was purified by silica gel column chromatography(n-hexane/ethyl acetate) to give the title compound (0.252 g) as asolid.

¹H-NMR (CDCl₃) δ: 0.11 (6H, s), 0.94 (9H, s), 3.81 (3H, s), 4.70 (2H,s), 5.91 (1H, br s), 6.92 (1H, t, J=2.4 Hz), 7.08 (2H, d, J=8.5 Hz),7.26 (2H, d, J=8.5 Hz), 7.84 (1H, d, J=2.4 Hz), 7.97 (1H, d, J=2.4 Hz).

MS (m/z): 345 (M+H)⁺.

Step 2 {4-[(5-methoxypyridin-3-yl)amino]phenyl}methanol

To a solution of the compound (0.252 g) obtained in the above Step 1 inTHF (7.31 mL) was added 1N tetrabutylammonium fluoride THF solution(1.46 mL), and the mixture was stirred overnight at room temperature.The reaction mixture was concentrated under reduced pressure, brine wasadded to the residue, and the mixture was extracted three times withdichloromethane. The organic layer was dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (dichloromethane/ethylacetate) to give the title compound (0.117 g) as a solid.

¹H-NMR (CD₃OD) δ: 3.80 (3H, s), 4.54 (2H, s), 7.03 (1H, t, J=2.4 Hz),7.12 (2H, d, J=8.5 Hz), 7.29 (2H, d, J=8.5 Hz), 7.66 (1H, d, J=2.4 Hz),7.86 (1H, br s).

MS (m/z): 231 (M+H)⁺.

Reference Example D-96 4-(5-methoxy-6-phenoxypyridazin-3-yl)benzaldehyde

A mixture of the compound (100 mg) obtained in Reference Example D-28,phenol (37.8 mg), DMF (2.68 mL) and potassium carbonate (111 mg) wasstirred at 100° C. for 15 hr. The reaction solution was allowed to coolto room temperature. Water was added to the reaction solution, and themixture was extracted with ethyl acetate. The organic layer was washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (n-hexane/ethyl acetate) to give the titlecompound (63.9 mg) as a solid.

¹H-NMR (CDCl₃) δ: 4.12 (3H, s), 7.27-7.28 (2H, m), 7.33 (1H, s),7.41-7.44 (2H, m), 8.00-8.01 (2H, m), 8.04-8.05 (1H, m), 8.18-8.20 (2H,m), 10.09 (1H, s).

MS (m/z): 307 (M+H)⁺.

Reference Example E-15-({[(1S,3R)-3-(methylamino)cyclopentyl]amino}methyl)-1H-indole-2-carbonitrileStep 1 tert-butyl2-cyano-5-[({(1S,3R)-3-[methyl(2-nitrobenzene-1-sulfonyl)amino]cyclopentyl}amino)methyl]-1H-indole-1-carboxylate

To the compound (1.11 g) obtained in Step 4 of Reference Example A-3,tert-butyl 5-(bromomethyl)-2-cyano-indole-1-carboxylate (1.24 g)produced according to the method described in a literature (cancer cell2015, 27, 589-602) and potassium carbonate (1.02 g) was added DMF (12mL), and the mixture was stirred at room temperature for 2 hr, andallowed to stand overnight. Ethyl acetate and saturated brine were addedto the reaction solution, and the mixture was subjected to liquidseparation. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(n-hexane/ethyl acetate, dichloromethane/methanol) to give the titlecompound (916 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.37-1.60 (3H, m), 1.70-1.88 (2H, m), 1.73 (9H, s),2.04-2.12 (1H, m), 2.89 (3H, s), 3.08-3.14 (1H, m), 3.82 (2H, s),4.30-4.39 (1H, m), 7.30 (1H, s), 7.44 (1H, d, J=9.1 Hz), 7.55 (1H, s),7.59-7.63 (1H, m), 7.66-7.73 (2H, m), 7.98-8.02 (1H, m), 8.15 (1H, d,J=9.1 Hz).

MS (m/z): 554 (M+H)⁺.

Step 25-({[(1S,3R)-3-(methylamino)cyclopentyl]amino}methyl)-1H-indole-2-carbonitrile

The compound (916 mg) obtained in the above Step 1 was dissolved in amixed solvent of THF (10 mL) and methanol (10 mL), and4-isopropylbenzenethiol (0.504 mL) and cesium carbonate (1.08 g) wereadded thereto, and the mixture was stirred at room temperature for 1 hr,and allowed to stand overnight. Ethyl acetate and saturated brine wereadded to the reaction solution, and the mixture was subjected to liquidseparation. The organic layer was washed with saturated brine, and theaqueous layer was extracted five times with ethyl acetate. The organiclayers were combined, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby amino silica gel column chromatography (n-hexane/ethyl acetate,followed by dichloromethane/methanol) to give the title compound (0.281g).

¹H-NMR (CDCl₃) δ: 1.38-1.44 (1H, m), 1.59-1.71 (2H, m), 1.86-1.93 (2H,m), 2.15-2.22 (1H, m), 2.46 (3H, s), 3.07-3.14 (1H, m), 3.22-3.28 (1H,m), 3.78-3.86 (2H, m), 7.05 (1H, s), 7.17 (2H, s), 7.51 (1H, s).

Reference Example E-2(1R,3S,4S)—N¹-{[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}-4-methoxy-N³-methylcyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl[(1S,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-methoxycyclopentyl]methylcarbamate

To a suspension of the compound (0.460 g) obtained in Step 3 ofReference Example A-17 and the compound (0.525 g) obtained in ReferenceExample D-56 in dichloromethane (16 mL) were added successively sodiumtriacetoxyborohydride (1.35 g) and acetic acid (0.320 mL), and themixture was stirred at room temperature for 17 hr. Water (25 mL) wasadded to the reaction mixture, and the mixture was extracted twice (80mL, 50 mL) with a mixed solvent of dichloromethane/methanol (9/1). Theorganic layers were combined, and dried over anhydrous sodium sulfate.The solvent was evaporated under reduced pressure, and the obtainedresidue was purified by silica gel column chromatography(dichloromethane/methanol) to give the title compound (0.733 g) as asolid.

¹H-NMR (CDCl₃) δ: 1.47 (9H, s), 1.53-1.65 (1H, m), 1.83-1.99 (2H, m),2.17-2.27 (1H, m), 2.84 (3H, s), 3.25-3.36 (1H, m), 3.31 (3H, s), 3.85(2H, s), 3.87-3.94 (1H, m), 3.95 (3H, s), 4.07 (3H, s), 4.20-4.29 (1H,m), 7.24 (1H, d, J=1.8 Hz), 7.38-7.43 (2H, m), 7.48-7.53 (2H, m), 7.94(1H, d, J=1.8 Hz).

MS (m/z): 472 (M+H)⁺.

Step 2(1R,3S,4S)—N¹-{[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}-4-methoxy-N³-methylcyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 11 ofReference Example A-1, using the compound obtained in the above Step 1.

MS (m/z): 372 (M+H)⁺.

Reference Example E-3(1S,3S,4S)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxycyclohexane-1,3-diamineStep 1 tert-butyl[(1S,2S,5S)-5-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-methoxycyclohexyl]carbamate

To a mixture of the compound (0.143 g) obtained in Step 6 of ReferenceExample A-10, the compound (0.175 g) obtained in Reference Example D-26and chloroform (15 mL) was added tetraisopropyl orthotitanate (0.520 mL)at room temperature, and the mixture was stirred for 1.5 hr. Sodiumtriacetoxyborohydride (0.692 g) was added thereto, and the mixture wasstirred for 16 hr. Saturated aqueous sodium hydrogencarbonate solution(5.00 mL) and aqueous Rochelle salt solution (5.00 mL) were addedthereto, and the mixture was stirred for 2 hr, and the reaction mixturewas extracted with dichloromethane. The organic layer was dried overanhydrous sodium sulfate, the solvent was evaporated under reducedpressure, and the obtained residue was subjected to silica gel columnchromatography (ethyl acetate/methanol) using amino silica gel column asa charge column to give the title compound (0.179 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.22-1.66 (4H, m), 1.47 (9H, s), 1.92-2.24 (3H, m),2.81-2.90 (1H, m), 3.13-3.22 (1H, m), 3.38 (3H, s), 3.62-3.70 (1H, m),3.75-3.80 (1H, m), 3.88-3.94 (1H, m), 4.02 (3H, s), 4.23 (3H, s),5.96-6.22 (1H, m), 7.13 (1H, s), 7.43-7.48 (2H, m), 7.92-7.97 (2H, m).

MS (m/z): 473 (M+H)⁺.

Step 2(1S,3S,4S)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxycyclohexane-1,3-diamine

The title compound was obtained in the same manner as in Step 4 of A-3,using the above Step 1. 1H-NMR (CDCl₃) δ: 1.02-1.70 (6H, m), 1.98-2.06(1H, m), 2.10-2.19 (2H, m), 2.58-2.70 (2H, m), 2.75-2.85 (1H, m), 3.40(3H, s), 3.88 (2H, s), 4.02 (3H, s), 4.23 (3H, s), 7.13 (1H, s),7.41-7.47 (2H, m), 7.92-7.98 (2H, m).

MS (m/z): 373 (M+H)⁺.

Reference Example E-4 tert-butyl[(1S,2R,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-methoxycyclopentyl]methylcarbamate

The title compound was obtained in the same manner as in Step 1 ofReference Example E-2, using the compound obtained in Step 8 ofReference Example A-16 and the compound obtained in Reference ExampleD-26.

¹H-NMR (CDCl₃) δ: 1.47 (9H, s), 1.58-1.62 (1H, m), 1.79-1.88 (1H, m),2.08-2.18 (2H, m), 2.93 (3H, s), 3.10-3.17 (1H, m), 3.28 (3H, s),3.73-3.79 (1H, m), 3.82-3.90 (2H, m), 4.01-4.02 (1H, m), 4.02 (3H, s),4.23 (3H, s), 7.13 (1H, s), 7.46 (2H, d, J=8.0 Hz), 7.95 (2H, d, J=8.0Hz).

Example 15-({[(1R,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrilehydrochloride Step 1 tert-butyl2-cyano-5-({[(1R,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1H-indole-1-carboxylate

The compound (160 mg) obtained in Step 2 of Reference Example C-3 andtert-butyl 5-(bromomethyl)-2-cyano-indole-1-carboxylate (117 mg)produced according to the method described in a literature (cancer cell2015, 27, 589-602.) were dissolved in DMF (2 mL), and potassiumcarbonate (121 mg) was added thereto, and the mixture was stirredovernight at room temperature. Ethyl acetate and saturated brine wereadded to the reaction solution, and the mixture was subjected to liquidseparation. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(dichloromethane/methanol) to give the title compound (133 mg) as asolid.

¹H-NMR (CDCl₃) δ: 1.52-1.66 (2H, m), 1.71 (9H, s), 1.88-1.95 (1H, m),2.11-2.18 (2H, m), 2.34-2.43 (1H, m), 3.38-3.45 (1H, m), 3.62 (2H, q,J=10.3 Hz), 3.91 (2H, s), 4.70-4.78 (1H, m), 5.39 (1H, d, J=6.7 Hz),7.11 (1H, s), 7.28 (1H, s), 7.50 (1H, dd, J=8.8, 1.5 Hz), 7.62 (1H, s),8.17 (1H, d, J=8.8 Hz), 8.45 (1H, s).

MS (m/z): 571 (M+H)⁺.

Step 25-({[(1R,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrile

The compound (133 mg) obtained in the above Step 1 was dissolved inacetonitrile (5 mL), and tin(IV) chloride (ca. 1.0 mol/L,dichloromethane solution) (2.3 mL) was added thereto, and the mixturewas stirred at room temperature for 1 hr. Ethyl acetate and saturatedaqueous sodium hydrogencarbonate solution were added to the reactionsolution, and the insoluble substance was removed by filtration throughCelite, and the filtrate was subjected to liquid separation. The aqueouslayer was extracted with ethyl acetate, and the organic layers werecombined, washed with saturated brine, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The obtained residuewas purified by silica gel column chromatography(dichloromethane/methanol) to give the title compound (76 mg) as asolid.

¹H-NMR (CD₃OD) δ: 1.53-1.62 (1H, m), 1.63-1.73 (1H, m), 1.97-2.09 (2H,m), 2.16-2.23 (1H, m), 2.25-2.33 (1H, m), 3.39-3.46 (1H, m), 3.83 (2H,q, J=10.7 Hz), 3.91 (2H, s), 4.68-4.75 (1H, m), 7.15 (1H, s), 7.38 (1H,dd, J=8.8, 1.5 Hz), 7.43 (1H, d, J=8.8 Hz), 7.50 (1H, s), 7.64 (1H, s),8.31 (1H, s).

MS (m/z): 471 (M+H)⁺.

Step 35-({[(1R,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrilehydrochloride

The compound (76 mg) obtained in the above Step 2 was dissolved inethanol (3 mL), and 1N hydrochloric acid (0.178 mL) was added thereto,and the solvent was evaporated under reduced pressure. Diethyl ether wasadded to the residue, and the solid was collected by filtration to givethe title compound (62 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.65-1.74 (1H, m), 1.77-1.87 (1H, m), 2.06-2.13 (1H,m), 2.17-2.30 (3H, m), 3.67-3.74 (1H, m), 4.10 (2H, q, J=11.2 Hz), 4.23(2H, t, J=5.5 Hz), 4.69-4.77 (1H, m), 7.45 (1H, d, J=1.8 Hz), 7.55 (2H,s), 7.74 (1H, s), 7.89 (1H, s), 8.37 (1H, br s), 8.45 (1H, s), 9.27 (2H,br s), 12.62 (1H, s).

MS (m/z): 471 (M+H)⁺.

Example 24-methyl-5-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1-[(1H-pyrazol-4-yl)methyl]-1H-indole-2-carbonitrilehydrochloride Step 14-methyl-5-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1-{[1-(triphenylmethyl)-1H-pyrazol-4-yl]methyl}-1H-indole-2-carbonitrile

A mixture of the compound (0.362 g) obtained in Step 2 of ReferenceExample D-2, the compound (0.320 g) obtained in Step 2 of ReferenceExample C-4, DIPEA (0.373 mL), acetic acid (0.245 mL) anddichloromethane (15 mL) was stirred under argon atmosphere at roomtemperature for 30 min. Sodium triacetoxyborohydride (0.468 g) was addedto the reaction solution, and the mixture was stirred overnight at roomtemperature. Saturated aqueous sodium hydrogencarbonate solution wasadded to the reaction solution, and the mixture was extracted withdichloromethane. The organic layer was dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue wassubjected to amino silica gel column chromatography(dichloromethane/methanol, n-hexane/ethyl acetate) to give the titlecompound (0.238 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.52-1.56 (3H, m), 1.89-1.94 (3H, m), 2.29-2.36 (1H,m), 2.55 (3H, s), 3.24-3.28 (4H, m), 3.62 (2H, q, J=10.1 Hz), 3.87 (2H,s), 5.28 (2H, s), 5.37 (1H, t, J=8.5 Hz), 7.09-7.12 (6H, m), 7.19 (2H,t, J=3.9 Hz), 7.28-7.32 (11H, m), 7.45 (1H, s), 7.50 (1H, s), 8.42 (1H,s).

Step 24-methyl-5-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1-[(1H-pyrazol-4-yl)methyl]-1H-indole-2-carbonitrile

A mixture of the compound (0.228 g) obtained in the above Step 1,hydrogen chloride (4 mol/L, 1,4-dioxane solution, 0.75 mL), methanol(2.5 mL) and dichloromethane (15 mL) was stirred under argon atmosphereat room temperature for 30 min. The reaction solution was concentratedunder reduced pressure, and the residue was subjected to amino silicagel column chromatography (dichloromethane/methanol) to give the titlecompound (0.139 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.56-1.69 (3H, m), 1.94-1.96 (3H, m), 2.28-2.35 (1H,m), 2.55 (3H, s), 3.21-3.30 (4H, m), 3.63 (2H, q, J=10.1 Hz), 3.87 (2H,s), 5.36-5.38 (3H, m), 7.22 (1H, s), 7.25-7.28 (1H, m), 7.35-7.37 (2H,m), 7.56 (2H, s), 8.42 (1H, s).

MS (m/z): 579 (M+H)⁺.

Step 34-methyl-5-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1-[(1H-pyrazol-4-yl)methyl]-1H-indole-2-carbonitrilehydrochloride

A mixture of the compound (0.129 g) obtained in the above Step 2, 1Nhydrochloric acid (0.210 mL) and ethanol (4.0 mL) was stirred at roomtemperature for 30 min. The reaction solution was concentrated underreduced pressure, and the residue was suspended in diethyl ether, andcollected by filtration to give the title compound (0.133 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.90-1.98 (3H, m), 2.13-2.15 (1H, m), 2.35-2.41 (1H,m), 2.50-2.51 (1H, m), 2.59 (3H, s), 3.27 (3H, s), 3.73-3.75 (1H, m),4.09 (2H, q, J=11.3 Hz), 4.26 (2H, s), 5.28-5.31 (1H, m), 5.44 (2H, s),7.55 (1H, d, J=8.5 Hz), 7.67 (1H, s), 7.74-7.76 (2H, m), 8.38 (1H, s),8.96 (1H, br s), 9.07 (1H, br s).

MS (m/z): 579 (M+H)⁺.

Example 35-({[(1S,3R)-3-{methyl[2-(2,2,2-trifluoroethyl)[1,3]thiazolo[5,4-d]pyrimidin-7-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrilehydrochloride Step 15-({[(1S,3R)-3-{methyl[2-(2,2,2-trifluoroethyl)[1,3]thiazolo[5,4-d]pyrimidin-7-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrile

A mixture of the compound (0.0790 g) obtained in Step 2 of ReferenceExample E-1,7-chloro-2-(2,2,2-trifluoroethyl)[1,3]thiazolo[5,4-d]pyrimidine (75.0mg) produced according to the method described in a literature (WO2016/195776), potassium carbonate (0.0610 g) and DMF (2 mL) was stirredunder argon atmosphere at room temperature for 1 hr. Water was added tothe reaction solution, and the mixture was extracted with ethylacetate/diethyl ether. The organic layer was dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue wassubjected to diol-modified silica gel column chromatography(dichloromethane/methanol) to give the title compound (0.0184 g) as asolid. 1H-NMR (CDCl₃) δ: 1.53-1.71 (3H, m), 1.90-2.04 (3H, m), 2.27-2.33(1H, m), 3.27 (1H, t, J=6.7 Hz), 3.42 (3H, s), 3.78-3.95 (4H, m), 5.88(1H, br s), 7.17 (1H, s), 7.39 (2H, s), 7.63 (1H, s), 8.41 (1H, s), 8.59(1H, br s).

MS (m/z): 486 (M+H)⁺.

Step 25-({[(1S,3R)-3-{methyl[2-(2,2,2-trifluoroethyl)[1,3]thiazolo[5,4-d]pyrimidin-7-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrilehydrochloride

The title compound was obtained in the same manner as in Step 3 ofExample 2, using the compound obtained in the above Step 1.

¹H-NMR (DMSO-D₆) δ: 1.01-1.35 (2H, m), 1.81-2.23 (4H, m), 3.25-3.78 (4H,m), 4.19-4.52 (4H, m), 5.61-5.89 (1H, m), 7.44-7.67 (3H, m), 7.85-7.92(1H, m), 8.41-8.48 (1H, m), 9.14 (1H, br s), 9.23 (1H, br s), 12.59 (1H,s).

MS (m/z): 486 (M+H)⁺.

Example 45-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[3,2-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-1H-indole-2-carbonitrile

The compound (83 mg) obtained in Step 2 of Reference Example E-1 and thecompound (78 mg) obtained in Step 4 of Reference Example B-6 weredissolved in 2-propanol (5 mL), and DIPEA (0.108 mL) was added thereto,and the mixture was stirred at 70° C. for 5 hr, and allowed to standovernight at room temperature. The reaction mixture was subjected tosilica gel column chromatography (diol silica gel,dichloromethane/methanol), and then purified by ChiralFlash (registeredtrademark, Daicel Corporation) IA (n-hexane/IPA) to give the titlecompound.

1H-NMR (CDCl₃) δ: 1.55-1.72 (2H, m), 1.90-2.02 (3H, m), 2.25-2.33 (1H,m), 3.26-3.33 (1H, m), 3.36 (3H, s), 3.68 (2H, q, J=10.1 Hz), 3.90 (2H,s), 5.27-5.35 (1H, m), 7.15 (1H, s), 7.32-7.37 (3H, m), 7.61 (1H, s),8.52 (1H, s), 10.19 (1H, br s).

MS (m/z): 485 (M+H)⁺.

Example 54-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]amino}cyclopentyl]amino}methyl)phenolhydrochloride Step 1(1R,3S)—N³-[(4-methoxyphenyl)methyl]-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]cyclopentane-1,3-diamine

The title compound (containing impurities, purity ca.80%) was obtainedin the same manner as in Step 1 of Example 2, using the compoundobtained in Step 2 of Reference Example C-6 and 4-anisaldehyde (CAS:1122-91-4).

¹H-NMR (CDCl₃) δ: 1.61-1.70 (2H, m), 1.88-1.98 (1H, m), 1.99-2.08 (2H,m), 2.30-2.39 (1H, m), 3.16-3.25 (1H, m), 3.27 (3H, s), 3.51 (2H, q,J=10.5 Hz), 3.73 (2H, s), 3.81 (3H, s), 4.84-4.94 (1H, m), 6.88 (2H, d,J=8.5 Hz), 7.25 (2H, d, J=8.5 Hz), 7.61 (1H, dd, J=8.5, 1.2 Hz), 7.84(1H, d, J=8.5 Hz), 7.89 (1H, s), 8.64 (1H, s).

MS (m/z): 445 (M+H)⁺.

Step 24-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]amino}cyclopentyl]amino)methyl)phenol

The compound (0.0920 g, containing impurities, purity ca.80%) obtainedin the above Step 1 was dissolved in dichloromethane (4.2 mL), and borontribromide (CAS: 10294-33-4) (1N dichloromethane solution, 2.6 mL) wasadded thereto, and the mixture was stirred at room temperature for 50min. The reaction solution was cooled to 0° C., and saturated aqueoussodium bicarbonate solution was added slowly thereto. The reactionmixture was stirred vigorously at 0° C. for 5 min, and extracted withdichloromethane/methanol (9/1), and the organic layer was dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography(dichloromethane/methanol) to give the title compound (0.0653 g) as asolid.

¹H-NMR (CDCl₃) δ: 1.93-2.29 (5H, m), 2.40-2.51 (1H, m), 3.34 (3H, s),3.38-3.46 (1H, m), 3.51 (2H, q, J=10.5 Hz), 3.81 (1H, d, J=12.8 Hz),3.87 (1H, d, J=12.8 Hz), 4.69-4.81 (1H, m), 6.81 (2H, d, J=8.5 Hz), 7.27(2H, d, J=8.5 Hz), 7.63 (1H, d, J=8.5 Hz), 7.84 (1H, d, J=8.5 Hz), 7.89(1H, s), 8.28 (1H, br s).

MS (m/z): 431 (M+H)⁺.

Step 34-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)quinazolin-4-yl]amino}cyclopentyl]amino}methyl)phenolhydrochloride

The title compound was obtained in the same manner as in Step 3 ofExample 2, using the compound obtained in the above Step 2.

1H-NMR (DMSO-D₆) δ: 1.96-2.25 (5H, m), 2.42-2.51 (1H, m), 3.51-3.62 (1H,m), 3.54 (3H, s), 3.90-4.09 (4H, m), 5.21-5.42 (1H, m), 6.81 (2H, d,J=8.5 Hz), 7.39 (2H, d, J=8.5 Hz), 7.91 (1H, d, J=8.5 Hz), 8.01 (1H, d,J=8.5 Hz), 8.32 (1H, br s), 8.84 (1H, s), 9.34-9.46 (1H, m), 9.57-9.70(1H, m), 9.74 (1H, br s).

MS (m/z): 431 (M+H)⁺.

Example 64-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-N-{3-[(prop-2-enoyl)amino]propyl}benzamidehydrochloride Step 1 tert-butyl{3-[4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)benzamide]propyl}carbamate

The title compound was obtained in the same manner as in Step 1 ofExample 2, using the compound obtained in Step 2 of Reference ExampleC-4 and the compound obtained in Step 1 of Reference Example D-85.

¹H-NMR (CDCl₃) δ: 1.46 (9H, s), 1.50-1.73 (5H, m), 1.85-1.99 (3H, m),2.24-2.30 (1H, m), 3.20-3.28 (3H, m), 3.31 (3H, s), 3.49-3.54 (2H, m),3.63 (2H, q, J=10.3 Hz), 3.84 (2H, t, J=14.0 Hz), 4.90-4.95 (1H, m),5.30-5.38 (1H, m), 7.28 (1H, br s), 7.34 (1H, s), 7.41 (2H, d, J=8.5Hz), 7.83 (2H, d, J=8.5 Hz), 8.41 (1H, s).

MS (m/z): 621 (M+H)⁺.

Step 2N-(3-aminopropyl)-4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)benzamidehydrochloride

The title compound (608 mg) was obtained in the same manner as in Step11 of Reference Example A-1, using the compound obtained in the aboveStep 1.

¹H-NMR (DMSO-D₆) δ: 1.81-1.93 (3H, m), 2.04-2.23 (4H, m), 2.34-2.41 (1H,m), 2.79-2.86 (2H, m), 3.32-3.38 (2H, m), 3.43 (3H, s), 4.13-4.22 (4H,m), 5.28-5.37 (1H, m), 7.74 (2H, d, J=7.9 Hz), 7.89 (1H, s), 7.95 (2H,d, J=7.9 Hz), 8.11 (3H, br s), 8.65 (1H, s), 8.90 (1H, t, J=5.8 Hz),9.89-9.98 (1H, m), 10.09-10.19 (1H, m).

MS (m/z): 521 (M+H)⁺.

Step 34-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-N-{3-[(prop-2-enoyl)amino]propyl}benzamidehydrochloride

To the compound (250 mg) obtained in the above Step 2 were addeddichloromethane (5 mL) and DIPEA (0.435 mL). Acryloyl chloride (0.022mL) was added thereto under ice-cooling, and the mixture was stirred at0° C. for 1 hr. Water and dichloromethane were added to the reactionsolution, and the mixture was subjected to liquid separation. Theorganic layer was dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (dichloromethane/methanol) to give a free form (37mg) of the title compound as a solid. This was dissolved in ethanol (3mL), and 1N hydrochloric acid (0.064 mL) was added thereto, and thesolvent was evaporated under reduced pressure. Ether was added to theresidue, and the solid was collected by filtration to give the titlecompound (39 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.66-1.73 (2H, m), 1.87-2.10 (5H, m), 2.31-2.38 (1H,m), 3.17-3.22 (2H, m), 3.27-3.31 (5H, m), 3.59-3.65 (1H, m), 4.09 (2H,q, J=11.1 Hz), 4.23 (2H, t, J=6.1 Hz), 5.23-5.31 (1H, m), 5.58 (1H, dd,J=10.3, 2.1 Hz), 6.07 (1H, dd, J=17.0, 2.1 Hz), 6.22 (1H, dd, J=17.0,10.3 Hz), 7.66 (2H, d, J=7.9 Hz), 7.72 (1H, s), 7.92 (2H, d, J=7.9 Hz),8.20 (1H, t, J=5.5 Hz), 8.38 (1H, s), 8.59 (1H, t, J=5.8 Hz), 9.30-9.38(1H, m), 9.43-9.51 (1H, m).

MS (m/z): 575 (M+H)⁺.

Example 7(1R,3S)—N³-{[4-(benzylamino)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 tert-butyl benzyl[4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]carbamate

The title compound was obtained in the same manner as in Step 1 ofExample 2, using the compound obtained in Step 2 of Reference ExampleC-4 and the compound obtained in Step 3 of Reference Example D-89.

¹H-NMR (CDCl₃) δ: 1.42 (9H, s), 1.55-1.68 (2H, m), 1.85-1.96 (3H, m),2.21-2.28 (1H, m), 3.18-3.25 (1H, m), 3.30 (3H, s), 3.63 (2H, q, J=10.2Hz), 3.74 (2H, s), 4.82 (2H, s), 5.28-5.35 (1H, m), 7.08-7.16 (2H, m),7.20-7.26 (5H, m), 7.27-7.35 (3H, m), 8.41 (1H, s).

MS (m/z): 626 (M+H)⁺.

Step 2(1R,3S)—N³-{[4-(benzylamino)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine

To a solution of the compound (397 mg) obtained in the above Step 1 indichloromethane (1.5 mL) was added trifluoroacetic acid (CAS: 76-05-1)(1.5 mL) at room temperature. The mixture was stirred at roomtemperature for 15 hr, and the reaction solution was basified withsaturated aqueous sodium hydrogencarbonate solution, and extracted withchloroform. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure,and the residue was subjected to reversed phase high-performance liquidchromatography (water/acetonitrile/0.1% formic acid) to give the titlecompound (301 mg) as a solid.

¹H-NMR (CDCl₃) δ: 1.46-1.62 (3H, m), 1.85-1.98 (3H, m), 2.21-2.29 (1H,m), 3.19-3.27 (1H, m), 3.30 (3H, s), 3.62 (2H, q, J=10.1 Hz), 3.67 (2H,s), 4.04 (1H, br s), 4.32 (2H, s), 5.27-5.36 (1H, m), 6.61 (2H, d, J=8.6Hz), 7.12 (2H, d, J=8.6 Hz), 7.27-7.30 (1H, m), 7.32-7.39 (5H, m), 8.41(1H, s).

Step 3(1R,3S)—N³-{[4-(benzylamino)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 3 ofExample 2, using the compound obtained in the above Step 2.

1H-NMR (DMSO-D₆) δ: 1.81-2.10 (5H, m), 2.28-2.35 (1H, m), 3.25 (3H, s),3.55 (1H, br s), 3.94 (2H, s), 4.08 (2H, q, J=11.0 Hz), 4.30 (2H, s),5.19-5.28 (1H, m), 6.56 (1H, br s), 6.60 (2H, d, J=8.6 Hz), 7.17-7.24(3H, m), 7.29-7.36 (4H, m), 7.70 (1H, s), 8.36 (1H, s), 8.79-9.04 (2H,m).

MS (m/z): 526 (M+H)⁺.

Example 8(1R,3S)—N³-({4-[(5-methoxypyridin-3-yl)amino]phenyl}methyl)-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1 4-[(5-methoxypyridin-3-yl)amino]benzaldehyde

To a solution of the compound (0.117 g) obtained in Step 2 of ReferenceExample D-94 in THF (5.07 mL) was added manganese dioxide (0.441 g), andthe mixture was stirred at room temperature for 5 hr, and allowed tostand overnight. Additional manganese dioxide (0.441 g) was addedthereto, and the mixture was stirred at 45° C. for 2 hr. The reactionmixture was allowed to cool to room temperature, and filtered throughCelite, and the filtrate was concentrated under reduced pressure. Theobtained crude product of the title compound was directly used in thenext step.

Step 2(1R,3S)—N³-({4-[(5-methoxypyridin-3-yl)amino]phenyl}methyl)-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 1 and Step3 of Example 2, using the compound obtained in the above Step 1 and thecompound obtained in Step 2 of Reference Example C-4.

¹H-NMR (DMSO-D₆) δ: 1.86-1.89 (1H, br m), 2.00-2.08 (4H, m), 2.33-2.38(1H, m), 3.30 (3H, s), 3.61 (1H, br s), 3.91 (3H, s), 4.06-4.12 (4H, m),5.28 (1H, br s), 7.31 (2H, d, J=8.5 Hz), 7.46 (1H, s), 7.58 (2H, d,J=8.5 Hz), 7.75 (1H, s), 8.05-8.07 (2H, m), 8.40 (1H, s), 9.37-9.52 (3H,br m).

MS (m/z): 543 (M+H)⁺.

Example 9(1R,3S)—N¹-[6-(cyclopropylmethyl)thieno[2,3-d]pyrimidin-4-yl]-N³-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N¹-methylcyclopentane-1,3-diaminehydrochloride

The compound (205 mg) obtained in Reference Example C-29 was dissolvedin dichloromethane (5 mL), and hydrogen chloride (4 mol/L, 1,4-dioxanesolution, 5 mL) was added thereto, and the mixture was stirred at roomtemperature for 1 hr. The solvent was evaporated under reduced pressure.Dichloromethane (5 mL) and DIPEA (0.266 mL) were added to the obtainedresidue. The compound (119 mg) obtained in Reference Example D-65,sodium triacetoxyborohydride (324 mg) and acetic acid (0.132 mL) wereadded thereto, and the mixture was stirred at room temperature for 1 hr.Dichloromethane and saturated aqueous sodium hydrogencarbonate solutionwere added thereto, and the mixture was subjected to liquid separation,and the aqueous layer was extracted with dichloromethane. The organiclayers were combined, washed with saturated aqueous sodiumhydrogencarbonate solution, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (dichloromethane/methanol, followedby ethyl acetate/methanol) to give a free form (40 mg) of the titlecompound. This was dissolved in ethanol (3 mL), and 1N hydrochloric acid(0.240 mL) was added thereto, and the solvent was evaporated underreduced pressure. Diethyl ether was added to the residue, and the solidwas collected by filtration to give the title compound (23 mg) as asolid.

¹H-NMR (DMSO-D₆) δ: 0.29-0.32 (2H, m), 0.54-0.59 (2H, m), 1.02-1.09 (1H,m), 1.86-1.91 (1H, m), 2.06-2.17 (4H, m), 2.34-2.41 (1H, m), 2.82 (2H,d, J=7.3 Hz), 3.36 (3H, s), 3.59-3.65 (1H, m), 4.01 (3H, s), 4.24 (2H,br s), 5.23-5.32 (1H, m), 7.51 (1H, s), 7.80 (2H, d, J=7.9 Hz), 7.96(2H, d, J=7.9 Hz), 8.14 (1H, s), 8.48 (1H, s), 8.54 (1H, s), 8.76 (1H,s), 9.74 (1H, br s), 9.94 (1H, br s).

MS (m/z): 500 (M+H)⁺.

Example 10N-(oxetan-3-yl)-4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)benzamidehydrochloride Step 1 methyl4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)benzoate

The title compound was obtained in the same manner as in Step 1 ofExample 2, using the compound obtained in Step 2 of Reference ExampleC-2 and methyl 4-formylbenzoate (CAS: 1571-08-0).

¹H-NMR (CDCl₃) δ: 1.19-1.43 (2H, m), 1.49-1.72 (2H, m), 1.79-2.00 (3H,m), 2.28 (1H, br s), 2.99 (1H, br s), 3.56 (2H, q, J=10.0 Hz), 3.88 (3H,s), 3.91-4.06 (2H, m), 4.26 (1H, br s), 6.60-6.79 (1H, m), 7.06 (1H, brs), 7.50 (2H, d, J=8.0 Hz), 7.98 (2H, d, J=8.0 Hz), 8.42 (1H, s).

MS (m/z): 479 (M+H)⁺.

Step 2 methyl4-({(tert-butoxycarbonyl)[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)benzoate

The title compound was obtained in the same manner as in Step 7 ofReference Example A-16, using the compound obtained in the above Step 1.

¹H-NMR (CDCl₃) δ: 1.02-1.12 (1H, m), 1.26-1.56 (12H, m), 1.72-1.86 (2H,m), 2.11 (1H, d, J=11.7 Hz), 2.22 (1H, d, J=11.0 Hz), 3.62 (2H, q,J=10.2 Hz), 3.91 (3H, s), 4.04-4.15 (1H, m), 4.25 (1H, br s), 4.42 (2H,br s), 4.88 (1H, d, J=7.4 Hz), 7.00 (1H, s), 7.30 (2H, d, J=8.0 Hz),7.98 (2H, d, J=8.0 Hz), 8.46 (1H, s).

MS (m/z): 579 (M+H)⁺.

Step 34-({(tert-butoxycarbonyl)[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)benzoicacid

The title compound was obtained in the same manner as in Step 9 ofReference Example A-1, using the compound obtained in the above Step 2.

¹H-NMR (DMSO-D₆) δ: 1.09-1.36 (9H, m), 1.35-1.55 (5H, m), 1.76 (1H, d,J=11.7 Hz), 1.86 (1H, d, J=11.7 Hz), 1.96 (1H, d, J=10.0 Hz), 4.04 (2H,q, J=11.7 Hz), 4.05-4.22 (2H, m), 4.42 (2H, br s), 7.34 (2H, d, J=8.0Hz), 7.61 (1H, s), 7.83 (1H, d, J=7.4 Hz), 7.89 (2H, d, J=8.0 Hz), 8.32(1H, s), 12.84 (1H, s).

MS (m/z): 565 (M+H)⁺.

Step 4 tert-butyl({4-[(oxetan-3-yl)carbamoyl]phenyl}methyl)[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]carbamate

The title compound was obtained in the same manner as in Step 3 ofReference Example C-32, using the compound obtained in the above Step 3and 3-oxetanamine.

¹H-NMR (CDCl₃) δ: 1.02-1.13 (1H, m), 1.24-1.58 (13H, m), 1.76 (1H, d,J=11.7 Hz), 1.84 (1H, d, J=9.8 Hz), 2.10 (1H, d, J=10.4 Hz), 2.20 (1H,d, J=11.0 Hz), 3.56-3.66 (2H, m), 4.24 (1H, br s), 4.41 (2H, br s), 4.60(2H, td, J=6.7, 2.5 Hz), 4.94-5.01 (1H, m), 5.01 (2H, t, J=7.1 Hz), 5.23(1H, dt, J=6.7, 6.1 Hz), 6.69 (1H, d, J=6.1 Hz), 7.02 (1H, s), 7.30 (2H,d, J=8.0 Hz), 7.72 (2H, br s), 8.45 (1H, s).

MS (m/z): 620 (M+H)⁺.

Step 5N-(oxetan-3-yl)-4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)benzamide

To a solution of the compound (47.9 mg) obtained in the above Step 4 indichloromethane (0.4 mL) was added trifluoroacetic acid (CAS: 76-05-1)(0.2 mL) at room temperature. The mixture was stirred at roomtemperature for 15 hr, and the reaction solution was basified withsaturated aqueous sodium hydrogencarbonate solution, and extracted withchloroform. The organic layer was washed with saturated brine, driedover anhydrous sodium sulfate, and concentrated under reduced pressure,and the residue was subjected to reversed phase high-performance liquidchromatography (water/acetonitrile/0.1% formic acid) to give the titlecompound (34.1 mg) as a solid.

¹H-NMR (CD₃OD) δ: 1.39-1.63 (4H, m), 2.00-2.10 (2H, m), 2.22-2.28 (1H,m), 2.58-2.65 (1H, m), 3.35 (1H, s), 3.67 (1H, br s), 3.76-3.91 (4H, m),4.25 (1H, br s), 4.33 (2H, s), 4.48-4.61 (2H, m), 7.53 (1H, s), 7.66(2H, d, J=8.0 Hz), 8.17 (2H, d, J=8.0 Hz), 8.33 (1H, s), 8.48 (2H, brs).

Step 6N-(oxetan-3-yl)-4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)benzamidehydrochloride

The title compound was obtained in the same manner as in Step 3 ofExample 2, using the compound obtained in the above Step 5.

¹H-NMR (DMSO-D₆) δ: 1.21-1.50 (4H, m), 1.79-1.93 (2H, m), 2.08 (1H, brs), 2.32-2.39 (1H, m), 2.54 (1H, s), 2.88 (1H, br s), 3.47-3.53 (1H, m),3.63-3.73 (2H, m), 4.03-4.16 (4H, m), 4.35-4.47 (2H, m), 7.63-7.70 (3H,m), 8.00 (1H, br s), 8.11 (2H, d, J=8.0 Hz), 8.18 (1H, s), 8.32 (1H, s).

Example 115-[4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]pyridin-2(1H)-onehydrochloride Step 1(1R,3S)—N³-[(4-iodophenyl)methyl]-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine

The title compound was obtained in the same manner as in Step 1 ofExample 2, using the compound obtained in Step 2 of Reference ExampleC-4 and 4-iodobenzaldehyde (CAS: 15164-44-0).

¹H-NMR (CDCl₃) δ: 1.58-1.73 (2H, m), 1.88-1.97 (3H, m), 2.23-2.30 (1H,m), 3.22-3.28 (1H, m), 3.30 (3H, s), 3.63 (2H, q, J=10.4 Hz), 3.73-3.80(2H, m), 5.04 (1H, br s), 5.27-5.35 (1H, m), 7.10 (2H, d, J=8.0 Hz),7.33 (1H, s), 7.66 (2H, d, J=8.0 Hz), 8.40 (1H, s).

Step 25-[4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]pyridin-2(1H)-one

The compound (0.110 g) obtained in the above Step 1 was dissolved in1,4-dioxane (2.00 mL), and2-hydroxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (CAS:1054483-78-1) (0.0600 g),[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloridedichloromethane adduct (0.0295 g) and saturated aqueous sodiumbicarbonate solution (1.00 mL) were added thereto, and the mixture wasstirred with heating in a microwave reactor at 110° C. for 10 min. Thereaction mixture was allowed to cool to room temperature, water wasadded thereto, and the mixture was extracted twice with dichloromethane.The organic layer was concentrated under reduced pressure, and theresidue was purified by amino silica gel chromatography(dichloromethane/ethyl acetate/methanol) to give the title compound(0.0828 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.22 (1H, dd, J=13.5, 6.7 Hz), 1.52-1.55 (1H, br m),1.65-1.66 (1H, br m), 1.96 (3H, d, J=8.6 Hz), 2.26-2.33 (1H, m),3.23-3.30 (4H, m), 3.63 (2H, q, J=10.0 Hz), 3.83 (2H, s), 5.30-5.35 (1H,br m), 6.70 (1H, d, J=9.2 Hz), 7.35-7.39 (5H, br m), 7.61 (1H, s), 7.78(1H, d, J=9.2 Hz), 8.41-8.42 (1H, br m).

MS (m/z): 514 (M+H)⁺.

Step 35-[4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]pyridin-2(1H)-onehydrochloride

The compound (0.0828 g) obtained in the above Step 2 was dissolved inethanol (3.00 mL), and 1N hydrochloric acid (0.161 mL) was addedthereto, and the solvent was evaporated under reduced pressure. Diethylether was added to the residue, and the solid was collected byfiltration to give the title compound (0.0505 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.87-2.11 (5H, m), 2.33-2.39 (1H, m), 3.28 (3H, s),3.62 (1H, s), 4.09 (2H, q, J=11.2 Hz), 4.18 (2H, t, J=5.5 Hz), 5.27 (1H,t, J=7.9 Hz), 6.46 (1H, d, J=9.8 Hz), 7.59 (2H, d, J=7.9 Hz), 7.66-7.68(2H, m), 7.72 (1H, s), 7.79 (1H, d, J=2.7 Hz), 7.89 (1H, dd, J=9.8, 2.7Hz), 8.37 (1H, s), 9.22 (1H, br s), 9.33 (1H, br s).

Example 12(1R,3S)—N¹-methyl-N³-({4-[(1-methyl-1H-pyrazol-4-yl)amino]phenyl}methyl)-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine

A mixture of the compound (50.0 mg) obtained in Step 1 of Example 11,1-methyl-1H-pyrazol-4-amine (CAS: 69843-13-6) (16.2 mg), cesiumcarbonate (89.5 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene(15.9 mg), palladium(II) acetate (4.8 mg) and 1,4-dioxane (1.0 mL) wasstirred with heating under nitrogen atmosphere in a microwave reactor at110° C. for 1 hr. Water was added to the reaction solution, and themixture was extracted with chloroform. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure, and the residue was subjected to reversed phasehigh-performance liquid chromatography (water/acetonitrile/0.1% formicacid) to give the title compound (9.86 mg) as a solid.

¹H-NMR (CD₃OD) δ: 1.92-2.13 (4H, m), 2.19-2.29 (1H, m), 2.47-2.55 (1H,m), 2.65 (1H, s), 3.37 (3H, s), 3.65-3.74 (1H, m), 3.87 (3H, s), 3.90(2H, q, J=10.7 Hz), 4.11 (2H, s), 5.24-5.33 (1H, m), 6.85 (2H, d, J=8.6Hz), 7.27 (2H, d, J=8.6 Hz), 7.37 (1H, s), 7.56 (1H, s), 7.63 (1H, s),8.26 (1H, s), 8.48 (1H, br s).

MS (m/z): 516 (M+H)⁺.

Example 13(1R,3S)—N³-{[4-(6′-chloro[2,3′-bipyridine]-5-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine

The compound (0.269 g) obtained in Step 1 of Example 11 was dissolved inethylene glycol dimethyl ether (2.28 mL), and6-chloro-3-pyridinylboronic acid (CAS: 444120-91-6) (0.143 g),bis(triphenylphosphine)palladium(II) dichloride dichloromethane adduct(0.0320 g), potassium carbonate (0.189 g) and water (0.228 mL) wereadded thereto, and the mixture was stirred under nitrogen atmosphere at60° C. for 4 hr. Water was added to the reaction solution, and themixture was extracted twice with dichloromethane. The organic layer wasconcentrated under reduced pressure, and the residue was purified bysilica gel chromatography (dichloromethane/methanol) to give the titlecompound (0.0101 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.52-1.67 (3H, br m), 1.94-1.97 (2H, br m), 2.26-2.33(1H, m), 3.24-3.29 (1H, m), 3.31 (3H, s), 3.62 (2H, q, J=10.2 Hz), 3.85(2H, s), 5.35 (1H, q, J=8.3 Hz), 7.34 (1H, s), 7.39 (1H, d, J=8.5 Hz),7.44-7.48 (3H, m), 7.51-7.56 (3H, m), 7.64-7.69 (1H, m), 7.83 (1H, dd,J=8.5, 2.7 Hz), 8.41 (1H, s), 8.60 (1H, s).

MS (m/z):610 (M+H)⁺.

Example 14(1R,3S)—N³-{[4-(furo[3,2-b]pyridin-6-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 2 and Step3 of Example 11, using the compound obtained in Step 1 of Example 11 and6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)furo[3,2-b]pyridine (CAS:1188539-34-5).

1H-NMR (DMSO-D₆) δ: 1.88-2.13 (4H, m), 2.33-2.42 (2H, m), 3.29 (3H, s),3.65 (1H, br s), 4.09 (2H, q, J=11.0 Hz), 4.25 (2H, t, J=6.4 Hz),5.24-5.33 (1H, m), 7.21 (1H, dd, J=2.4, 1.2 Hz), 7.71 (2H, d, J=7.9 Hz),7.73 (1H, s), 7.93 (2H, d, J=7.9 Hz), 8.38 (1H, s), 8.39 (1H, d, J=1.8Hz), 8.42 (1H, dd, J=1.8, 1.2 Hz), 8.92 (1H, d, J=1.8 Hz), 9.26 (1H, brs), 9.38 (1H, br s).

MS (m/z):538 (M+H)⁺.

Example 15(1R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-olhydrochloride (15A)(1S,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-olhydrochloride (15B) Step 1(1R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-ol(racemate)

The title compound (390 mg) was obtained in the same manner as in Step 1of Example 2, using the compound obtained in Step 2 of Reference ExampleC-18 and the compound obtained in Reference Example D-65.

¹H-NMR (DMSO-D₆) δ: 0.95-1.11 (2H, m), 1.25-1.28 (1H, m), 2.10-2.16 (3H,m), 2.52-2.55 (1H, m), 3.48-3.50 (1H, m), 3.79 (2H, s), 3.91 (3H, s),4.00-4.10 (3H, m), 4.71 (1H, d, J=4.9 Hz), 7.47 (2H, d, J=8.3 Hz),7.61-7.61 (1H, m), 7.65 (1H, s), 7.70 (2H, d, J=8.3 Hz), 7.85-7.87 (1H,m), 8.27-8.27 (1H, m), 8.31 (1H, s), 8.48 (1H, s).

MS (m/z): 544 (M+H)⁺.

Step 2(1R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-ol(a free form of 15A)(1S,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-ol(a free form of 15B)

The compound (390 mg) obtained in the above Step 1 was subjected tooptical resolution with chiral high-performance liquid chromatography(CHIRALPAK (registered trademark, Daicel Corporation) IA, mobile phase:n-hexane/2-propanol) to give a free form (an earlier eluted component,150 mg) of 15A, and a free form (a later eluted component, 185 mg) of15B, respectively as a solid.

the free form of 15A (earlier eluted component)

¹H-NMR (DMSO-D₆) δ: 0.95-1.29 (3H, m), 2.04-2.19 (4H, m), 2.47-2.57 (1H,m), 3.47-3.51 (1H, m), 3.78-3.79 (2H, m), 3.91 (3H, s), 3.99-4.10 (3H,m), 4.71-4.73 (1H, m), 7.47 (2H, d, J=8.3 Hz), 7.61-7.61 (1H, m), 7.66(1H, s), 7.70 (2H, d, J=8.3 Hz), 7.87 (1H, d, J=8.0 Hz), 8.27-8.28 (1H,m), 8.31 (1H, s), 8.48 (1H, s).

MS (m/z): 544 (M+H)⁺.

the free form of 15B (later eluted component)

¹H-NMR (DMSO-D₆) δ: 1.05-1.25 (3H, m), 2.12-2.19 (4H, m), 2.54-2.57 (1H,m), 3.51-3.54 (1H, m), 3.80 (2H, s), 3.92 (3H, s), 4.03-4.14 (3H, m),4.76-4.78 (1H, m), 7.48 (2H, d, J=8.0 Hz), 7.62-7.62 (1H, m), 7.69 (1H,s), 7.71 (2H, d, J=8.0 Hz), 7.91 (1H, d, J=7.4 Hz), 8.29 (1H, s), 8.33(1H, s), 8.49 (1H, s). separation condition (analysis) CHIRALPAK(registered trademark, Daicel Corporation) IA, size 0.46 cm×15 cm, flowrate 1.0 mL/min, mobile phase: n-hexane/2-propanol=20/80 to 0/100,temperature 26° C.

the free form of 15A retention time 3.0 min, the free form of 15Bretention time 3.6 min

separation condition (preparative) CHIRALPAK (registered trademark,Daicel Corporation) IA, size 2.5 cm×25 cm, flow rate 20 mL/min, mobilephase n-hexane/2-propanol=50/50, temperature 40° C.

the free form of 15A retention time 5.7 min, the free form of 15Bretention time 7.2 min

Step 3(1R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-olhydrochloride (15A)

A mixture of the free form of 15A (lower polar isomer, 149 mg) obtainedin the above Step 2, hydrochloric acid (1 mol/L, ethanol solution, 0.274mL) and ethanol (2 mL) was stirred, and concentrated under reducedpressure. Ethanol/diethyl ether solution was added to the residue, andthe resulting solid was collected by filtration to give the titlecompound (142 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.29-1.49 (3H, m), 2.10-2.13 (1H, m), 2.34-2.41 (2H,m), 3.24-3.31 (1H, m), 3.58-3.61 (2H, m), 3.91 (3H, s), 4.05-4.08 (2H,m), 4.19-4.24 (3H, m), 7.66-7.69 (4H, m), 7.84-7.86 (2H, m), 8.06-8.08(1H, m), 8.33-8.34 (2H, m), 8.54 (1H, s), 9.18-9.21 (2H, m).

MS (m/z):544 (M+H)⁺.

Step 4(1S,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-olhydrochloride (15B)

A mixture of the free form of 15B (125 mg) obtained in the above Step 9,hydrochloric acid (1 mol/L, ethanol solution, 0.23 mL) and ethanol (2mL) was stirred, and concentrated under reduced pressure.Ethanol/diethyl ether solution was added to the residue, and theresulting solid was collected by filtration to give the title compound(116 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.40-1.52 (3H, m), 2.11-2.13 (1H, m), 2.45-2.48 (2H,m), 3.28-3.31 (1H, m), 3.60-3.64 (2H, m), 3.95 (3H, s), 4.08-4.11 (2H,m), 4.25-4.27 (3H, m), 7.70 (1H, s), 7.73 (2H, d, J=8.5 Hz), 7.80 (1H,s), 7.89 (2H, d, J=8.5 Hz), 8.17-8.19 (1H, m), 8.38-8.39 (2H, m), 8.60(1H, s), 9.37-9.40 (2H, m).

MS (m/z):544 (M+H)⁺.

Example 16(1R,3S,5R)-3-[{[4-(5-methoxypyridin-3-yl)phenyl]methyl}(methyl)amino]-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexan-1-ol

A mixture of the compound (the free form of 15B) (0.0353 g) obtained inStep 2 of Example 15, dichloromethane (0.649 mL) and pyridine (0.0157mL) was ice-cooled, and methyl trifluoromethanesulfonate (0.0147 mL) wasadded thereto. The mixture was stirred at the same temperature for 2 hr,gradually warmed to room temperature, and stirred for 1 hr. The reactionmixture was ice-cooled, and additional pyridine (0.0261 mL) and methyltrifluoromethanesulfonate (0.0294 mL) were added thereto, and themixture was stirred at same temperature for 1 hr, and then at roomtemperature overnight. Saturated aqueous sodium bicarbonate solution wasadded to the reaction mixture, and the mixture was extracted three timeswith ethyl acetate. The organic layer was dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The residue waspurified by preparative TLC (dichloromethane/methanol) and amino silicagel preparative TLC (dichloromethane/ethyl acetate) to give the titlecompound (0.0028 g).

₁H-NMR (CD₃OD) δ: 0.88-0.91 (2H, m), 1.38-1.51 (3H, m), 1.59 (1H, s),2.23-2.32 (3H, m), 2.34 (3H, s), 2.85 (1H, t, J=11.6 Hz), 3.79 (2H, s),3.86 (2H, q, J=10.6 Hz), 3.95 (3H, s), 4.63 (1H, br s), 7.50 (1H, s),7.52 (2H, d, J=1.2 Hz), 7.63 (1H, s), 7.67 (2H, d, J=8.5 Hz), 8.22 (1H,s), 8.32 (1H, s), 8.39 (1H, s).

MS (m/z): 558 (M+H)⁺.

Example 17(1S,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamidehydrochloride (17A)(1R,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamidehydrochloride (17B) Step 1(1R,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamidehydrochloride (racemate)

To a mixture of the compound (0.0375 g) obtained in Step 2 of ReferenceExample C-17, the compound (0.0240 g) obtained in Reference Example D-65and chloroform (2.0 mL) was added tetraisopropyl orthotitanate (CAS:546-68-9) (0.083 mL) at room temperature. This reaction solution wasstirred at room temperature for 1.5 hr, sodium triacetoxyborohydride(purity 97%, 0.109 g) was added thereto, and the mixture was stirred atroom temperature for 6 hr, and allowed to stand for 16 hr. Saturatedaqueous sodium hydrogencarbonate solution (10 mL) and saturated aqueouspotassium sodium tartrate solution (10 mL) were added to this reactionsolution, and the mixture was extracted three times with dichloromethane(10 mL). The organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by aminosilica gel column chromatography (methanol/ethyl acetate) to give an oil(0.0457 g). To a mixture of the obtained oil (0.0457 g) and ethanol (3.0mL) was added 1N hydrogen chloride ethanol solution (0.076 mL) at roomtemperature. This reaction solution was stirred at room temperature for2.5 hr, and concentrated under reduced pressure. A suspension of theresidue in ethanol (0.05 mL) and diethyl ether (6.0 mL) was filtered,and the obtained solid was dried under reduced pressure at 40° C. for 2hr to give the title compound (0.0419 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.46-1.64 (3H, m), 1.91 (1H, d, J=12.2 Hz), 2.21(1H, d, J=12.2 Hz), 2.53-2.55 (1H, m), 2.84 (3H, s), 3.01-3.04 (1H, m),3.06 (3H, s), 3.93 (3H, s), 4.09 (2H, q, J=11.0 Hz), 4.26-4.28 (2H, m),4.34-4.36 (1H, m), 7.67 (1H, s), 7.69 (1H, dd, J=2.4, 1.8 Hz), 7.72 (2H,d, J=7.9 Hz), 7.87 (2H, d, J=7.9 Hz), 8.10 (1H, d, J=7.9 Hz), 8.33 (1H,d, J=2.4 Hz), 8.37 (1H, s), 8.55 (1H, d, J=1.8 Hz), 9.26 (1H, br s),9.34 (1H, br s).

Step 2(1S,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamide(a free form of 17A)(1R,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamide(a free form of 17B)

To a mixture of the compound (0.0339 g) obtained in the above Step 1 anddichloromethane (5.0 mL) was added saturated aqueous sodiumhydrogencarbonate solution (5.0 mL) at room temperature. This mixturewas stirred vigorously at room temperature for 1 hr, and extracted threetimes with dichloromethane (10 mL), and the organic layer was dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Theresidue was purified by chiral high-performance liquid chromatography(CHIRALPAK (registered trademark, Daicel Corporation) IA, mobile phase:2-propanol/n-hexane) to give a free form of 17A (an earlier elutedcomponent, 0.0137 g) and a free form of 17B (a later eluted component,0.0145 g), respectively as an oil.

the free form of 17A (earlier eluted component)

¹H-NMR (CDCl₃) δ: 1.31 (1H, q, J=11.5 Hz), 1.51 (1H, q, J=12.1 Hz), 1.65(1H, q, J=12.1 Hz), 1.92 (1H, br s), 2.12-2.18 (2H, m), 2.48-2.51 (1H,m), 2.85-2.89 (2H, m), 2.97 (3H, s), 3.11 (3H, s), 3.61 (2H, q, J=10.3Hz), 3.91-3.93 (2H, m), 3.93 (3H, s), 4.39-4.46 (1H, m), 5.92 (1H, d,J=8.5 Hz), 7.25 (1H, s), 7.35 (1H, dd, J=3.0, 1.8 Hz), 7.43 (2H, d,J=8.5 Hz), 7.54 (2H, d, J=8.5 Hz), 8.30 (1H, d, J=3.0 Hz), 8.45 (1H, d,J=1.8 Hz), 8.47 (1H, s).

the free form of 17B (later eluted component)

¹H-NMR (CDCl₃) δ: 1.31 (1H, q, J=11.5 Hz), 1.51 (1H, q, J=12.1 Hz), 1.65(1H, q, J=12.1 Hz), 1.90 (1H, br s), 2.12-2.18 (2H, m), 2.48-2.50 (1H,m), 2.84-2.89 (2H, m), 2.98 (3H, s), 3.11 (3H, s), 3.61 (2H, q, J=10.3Hz), 3.91-3.93 (2H, m), 3.93 (3H, s), 4.40-4.44 (1H, m), 5.91 (1H, d,J=8.5 Hz), 7.25 (1H, s), 7.36 (1H, dd, J=3.0, 1.8 Hz), 7.43 (2H, d,J=7.9 Hz), 7.54 (2H, d, J=7.9 Hz), 8.30 (1H, d, J=3.0 Hz), 8.45 (1H, d,J=1.8 Hz), 8.47 (1H, s).

separation condition (analysis) CHIRALPAK (registered trademark, DaicelCorporation) IA, size 0.46 cm×15 cm, flow rate 1.0 mL/min, mobile phasen-hexane/2-propanol=50/50 to 0/100, temperature 40° C.

the free form of 17A retention time 5.2 min, the free form of 17Bretention time 7.3 min

separation condition (preparative) CHIRALPAK (registered trademark,Daicel Corporation) IA, size 2.0 cm×25 cm, flow rate 10 mL/min, mobilephase n-hexane/2-propanol=25/75, temperature 20° C.

the free form of 17A retention time 14.0 min, the free form of 17Bretention time 19.8 min

Step 3(1S,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamidehydrochloride (17A)

To a mixture of the free form of 17A (0.0137 g) obtained in the aboveStep 2 and ethanol (5.0 mL) was added 1N hydrogen chloride ethanolsolution (0.030 mL) at room temperature. This reaction solution wasstirred at room temperature for 2.5 hr, and concentrated under reducedpressure. A suspension of the residue in ethanol (0.05 mL) and diethylether (6.0 mL) was filtered, and the obtained solid was dried underreduced pressure at 40° C. for 2 hr to give the title compound (0.0120g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.46-1.56 (3H, m), 1.91 (1H, d, J=13.4 Hz), 2.20(1H, d, J=13.4 Hz), 2.54 (1H, s), 2.84 (3H, s), 3.01-3.03 (1H, m), 3.06(3H, s), 3.93 (3H, s), 4.10 (2H, q, J=11.5 Hz), 4.29-4.31 (2H, m),4.33-4.36 (1H, m), 7.65 (1H, s), 7.69 (2H, d, J=7.6 Hz), 7.71-7.73 (1H,m), 7.88 (2H, d, J=7.6 Hz), 8.09 (1H, m), 8.35 (1H, d, J=2.4 Hz), 8.38(1H, s), 8.57 (1H, s), 9.09 (1H, br s), 9.16 (1H, br s).

MS (m/z): 599 (M+H)⁺.

Step 4(1R,3S,5R)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-N,N-dimethyl-5-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexane-1-carboxamidehydrochloride (17B)

To a mixture of the free form of 17B (0.0145 g) obtained in the aboveStep 2 and ethanol (5.0 mL) was added 1N hydrogen chloride ethanolsolution (0.030 mL) at room temperature. This reaction solution wasstirred at room temperature for 2.5 hr, and concentrated under reducedpressure. A suspension of the residue in ethanol (0.05 mL) and diethylether (6.0 mL) was filtered, and the obtained solid was dried underreduced pressure at 40° C. for 2 hr to give the title compound (0.0145g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.52-1.61 (3H, m), 1.90 (1H, d, J=12.1 Hz), 2.21(1H, d, J=12.1 Hz), 2.53-2.54 (1H, m), 2.84 (3H, s), 2.99-3.02 (1H, m),3.06 (3H, s), 3.94 (3H, s), 4.10 (2H, q, J=10.9 Hz), 4.27-4.29 (2H, m),4.35-4.36 (1H, m), 7.66-7.68 (1H, m), 7.73 (2H, d, J=8.2 Hz), 7.82 (1H,s), 7.90 (2H, d, J=8.2 Hz), 8.20 (1H, s), 8.38-8.39 (2H, m), 8.61 (1H,s), 9.33 (1H, br s), 9.41 (1H, br s).

MS (m/z): 599 (M+H)⁺.

Example 18(1R,3S,5R)-5-methoxy-N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride(1S,3R,5S)-5-methoxy-N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride Step 1(1R,3S,5R)-5-methoxy-N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine(racemate)

The title compound was obtained in the same manner as in Step 1 ofReference Example E-3, using the compound obtained in Step 2 ofReference Example C-18 and the compound obtained in Reference ExampleD-65.

1H-NMR (CDCl₃) δ: 1.25-1.39 (3H, m), 2.34-2.44 (3H, m), 2.83-2.88 (1H,m), 3.35 (3H, s), 3.39-3.44 (1H, m), 3.56-3.64 (2H, m), 3.89 (2H, s),3.92 (3H, s), 4.36 (1H, d, J=7.3 Hz), 5.64 (1H, br s), 7.03 (1H, s),7.36 (1H, s), 7.42 (2H, d, J=7.9 Hz), 7.55 (2H, d, J=7.9 Hz), 8.30 (1H,t, J=2.7 Hz), 8.45 (1H, t, J=2.1 Hz), 8.47 (1H, s).

MS (m/z): 558 (M+H)⁺.

Step 2(1R,3S,5R)-5-methoxy-N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride(1S,3R,5S)-5-methoxy-N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride

The compound (0.24 g) obtained in the above Step 1 was subjected tooptical resolution with chiral high-performance liquid chromatography(CHRALPAK (registered trademark, Daicel Corporation) IA, mobile phase:n-hexane/2-propanol/ethyl acetate/diethylamine) to give a free form of18A (an earlier eluted component, 69 mg) and a free form of 18B (a latereluted component, 51 mg).

The free form of 18A (0.069 g) was dissolved in methanol (3.00 mL), 1Nhydrochloric acid (0.124 mL) was added thereto, and the solvent wasevaporated under reduced pressure. Diethyl ether was added to theresidue, and the solid was collected by filtration to give one compound(18A) (0.069 g) of the title compounds, as a solid.

¹H-NMR (CD₃OD) δ: 1.37-1.59 (3H, m), 2.48-2.54 (1H, m), 2.64-2.70 (2H,m), 3.40-3.53 (2H, m), 3.44 (3H, s), 3.87 (2H, q, J=10.5 Hz), 3.98 (3H,s), 4.31-4.41 (3H, m), 7.54 (1H, s), 7.67 (2H, d, J=8.5 Hz), 7.73-7.75(1H, m), 7.82 (2H, d, J=8.5 Hz), 8.31 (1H, d, J=2.0 Hz), 8.37 (1H, s),8.46 (1H, br s).

MS (m/z): 558 (M+H)⁺.

The free form of 18B (0.051 g) was dissolved in methanol (3.00 mL), 1Nhydrochloric acid (0.092 mL) was added thereto, and the solvent wasevaporated under reduced pressure. Diethyl ether was added to theresidue, and the solid was collected by filtration to give the othercompound (18B) (0.054 g) of the title compounds, as a solid.

¹H-NMR (CD₃OD) δ: 1.34-1.53 (3H, m), 2.48-2.53 (1H, m), 2.64-2.70 (2H,m), 3.41-3.49 (2H, m), 3.44 (3H, s), 3.88 (2H, q, J=10.5 Hz), 3.97 (3H,s), 4.32-4.39 (3H, m), 7.52 (1H, s), 7.66 (2H, d, J=8.2 Hz), 7.73 (1H,br s), 7.83 (2H, d, J=8.2 Hz), 8.30 (1H, d, J=1.8 Hz), 8.37 (1H, s),8.45 (1H, br s).

MS (m/z): 558 (M+H)⁺.

separation condition (analysis) CHRALPAK (registered trademark, DaicelCorporation) IA, size 0.46 cm×25 cm, flow rate 1.0 mL/min, mobile phasen-hexane/2-propanol/ethyl acetate/diethylamine=50/30/20/0.1, temperature40° C. the free form of 18A retention time 7.2 min, the free form of 18Bretention time 9.8 min

separation condition (preparative) CHRALPAK (registered trademark,Daicel Corporation) IA, size 2.0 cm×25 cm, flow rate 5.7 mL/min, mobilephase n-hexane/2-propanol/ethyl acetate/diethylamine=50/30/20/0.1,temperature 40° C. the free form of 18A retention time 21 min, the freeform of 18B retention time 27 min

Example 19(1S,3R,5R)—N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-5-(1,3-oxazol-2-yl)-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride(1R,3S,5S)—N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-5-(1,3-oxazol-2-yl)-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride Step 1(1S,3R,5R)—N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-5-(1,3-oxazol-2-yl)-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine(racemate)

The title compound was obtained in the same manner as in Step 1 ofReference Example E-3, using the compound obtained in Step 6 ofReference Example C-32 and the compound obtained in Reference ExampleD-65.

¹H-NMR (CDCl₃) δ: 1.12-1.75 (4H, m), 2.42-2.63 (3H, m), 2.89-3.02 (1H,m), 3.05-3.19 (1H, m), 3.55-3.67 (2H, m), 3.88-3.98 (5H, m), 4.42-4.49(1H, m), 5.35 (1H, s), 6.96-7.00 (1H, m), 7.02 (1H, s), 7.35-7.38 (1H,m), 7.41-7.46 (2H, m), 7.52-7.58 (3H, m), 8.29-8.31 (1H, m), 8.45-8.47(1H, m), 8.48 (1H, s).

MS (m/z): 595 (M+H)⁺.

Step 2(1S,3R,5R)—N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-5-(1,3-oxazol-2-yl)-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride(1R,3S,5S)—N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-5-(1,3-oxazol-2-yl)-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride

The compound obtained in the above Step 1 (68 mg) was subjected tooptical resolution with chiral high-performance liquid chromatography(CHRALPAK (registered trademark, Daicel Corporation) IA, mobile phase:n-hexane/2-propanol) to give a free form of 19A (an earlier elutedcomponent, 22 mg) and a free form of 19B (a later eluted component, 23mg).

To a solution of the free form of 19A (22 mg) in ethanol (0.500 mL) wasadded 2N hydrochloric acid (0.0185 mL) at room temperature, and themixture was concentrated under reduced pressure. Ethanol (1.00 mL) wasadded thereto, and the mixture was azeotropically concentrated, and theresidue was solidified with diethyl ether to give one compound (19A) (23mg) of the title compounds, as a solid.

¹H-NMR (DMSO-D₆) δ: 1.53-1.77 (3H, m), 2.28-2.35 (1H, m), 2.57-2.63 (2H,m), 3.22-3.31 (1H, m), 3.92-3.94 (3H, m), 4.10 (2H, q, J=11.0 Hz),4.27-4.45 (3H, m), 7.18 (1H, s), 7.65-7.76 (4H, m), 7.89 (2H, d, J=8.6Hz), 8.09 (1H, s), 8.14 (1H, br s), 8.33-8.37 (1H, m), 8.40 (1H, s),8.55-8.59 (1H, m).

MS (m/z): 595 (M+H)⁺.

To a solution of the free form of 19B (23 mg) in ethanol (0.500 mL) wasadded 2N hydrochloric acid (0.0196 mL) at room temperature, and themixture was concentrated under reduced pressure. Ethanol (1.00 mL) wasadded thereto, and the mixture was azeotropically concentrated, and theresidue was solidified with diethyl ether to give the other compound(19B) (23 mg) of the title compounds, as a solid.

¹H-NMR (DMSO-D₆) δ: 1.54-1.77 (3H, m), 2.29-2.36 (1H, m), 2.54-2.67 (2H,m), 3.22-3.31 (1H, m), 3.93-3.93 (3H, m), 4.10 (2H, q, J=11.0 Hz),4.27-4.45 (3H, m), 7.18 (1H, s), 7.65-7.74 (4H, m), 7.89 (2H, d, J=8.0Hz), 8.09 (1H, s), 8.11-8.19 (1H, m), 8.35 (1H, br s), 8.40 (1H, s),8.54-8.58 (1H, m), 9.34 (2H, br s).

MS (m/z):595 (M+H)⁺.

separation condition (analysis) CHIRALPAK (registered trademark, DaicelCorporation) IA, size 0.46 cm×15 cm, flow rate 1.0 mL/min, mobile phase2-propanol 100%, temperature 40° C. the free form of 19A retention time5.4 min, the free form of 19B retention time 7.7 min

separation condition (preparative) CHIRALPAK (registered trademark,Daicel Corporation) IA, size 2 cm×25 cm, flow rate 10 mL/min, mobilephase n-hexane/2-propanol=20/80, temperature ca. 20° C.

the free form of 19A retention time 19.9 min, the free form of 19Bretention time 32.9 min

Example 20(1S,2R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentane-1,2-diolhydrochloride (20A)(1R,2S,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentane-1,2-diolhydrochloride (20B) Step 1(1S,2R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentane-1,2-diol

A mixture of the isomer A (193 mg) obtained in Step 7 of ReferenceExample C-33, dichloromethane (5 mL) and TFA (5 mL) was stirred at roomtemperature for 2 hr. The reaction solution was concentrated underreduced pressure to give an oil (283 mg). This was directly used in thenext step. A mixture of the above oil (283 mg), the compound (75.6 mg)obtained in Reference Example D-65, DIPEA (0.218 mL), dichloromethane(4.2 mL), sodium triacetoxyborohydride (265 mg) and acetic acid (0.143mL) was stirred at room temperature for 2.5 hr. The compound (25.0 mg)obtained in Reference Example D-65 was added again thereto, and themixture was stirred for additional 1.5 hr. Saturated aqueous sodiumhydrogencarbonate solution was added to the reaction solution, and themixture was extracted with ethyl acetate. The organic layer was washedwith saturated brine, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (ethyl acetate/methanol), followed bypreparative TLC (ethyl acetate/methanol) to give the title compound(30.3 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.94-2.07 (2H, m), 2.93-2.95 (1H, m), 3.37 (3H, s),3.82-3.87 (2H, m), 3.90 (3H, s), 3.95-3.97 (1H, m), 4.07-4.12 (3H, m),4.67-4.70 (1H, m), 4.97-4.99 (1H, m), 5.18-5.21 (1H, m), 7.50 (2H, d,J=8.0 Hz), 7.61-7.62 (1H, m), 7.72 (3H, d, J=8.0 Hz), 8.28 (1H, s), 8.31(1H, s), 8.49 (1H, s).

MS (m/z): 560 (M+H)⁺.

Step 2(1S,2R,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentane-1,2-diolhydrochloride (20A)

A mixture of the compound (28.3 mg) obtained in the above Step 1,hydrogen chloride (1 mol/L, ethanol solution, 0.0506 mL) and ethanol (1mL) was stirred at room temperature. The mixture was concentrated underreduced pressure. Ethanol/diethyl ether was added to the residue, andthe resulting solid was collected by filtration, and dried to give thetitle compound (25.0 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 2.22-2.33 (2H, m), 3.45 (3H, s), 3.94 (3H, s), 4.08(2H, q, J=11.0 Hz), 4.18-4.21 (1H, m), 4.24-4.27 (3H, m), 5.23-5.40 (2H,m), 5.92-5.95 (1H, m), 7.69 (2H, d, J=8.0 Hz), 7.74-7.75 (2H, m), 7.88(2H, d, J=8.0 Hz), 8.34-8.37 (2H, m), 8.57 (1H, s), 8.97 (1H, s), 9.16(1H, s).

MS (m/z): 560 (M+H)⁺.

Step 3(1R,2S,3R,5S)-3-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-5-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentane-1,2-diolhydrochloride (20B)

The title compound was obtained in the same manner as in the above Step1 and Step 2, using the isomer B obtained in Step 7 of Reference ExampleC-33.

1H-NMR (DMSO-D₆) δ: 1.85-1.91 (1H, m), 2.31-2.38 (1H, m), 3.30 (3H, s),3.92 (3H, s), 4.07-4.13 (3H, m), 4.31-4.35 (3H, m), 5.09 (1H, s), 5.26(2H, s), 7.67-7.75 (4H, m), 7.87 (2H, d, J=8.0 Hz), 8.32 (1H, d, J=3.1Hz), 8.38 (1H, s), 8.54 (1H, d, J=1.8 Hz), 9.53-9.60 (2H, m).

MS (m/z): 560 (M+H)+.

Example 21(1R,2S,4R)-2-{[2-amino-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)cyclopentan-1-olhydrochloride Step 1(1R,2S,4R)-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)-2-{[2-{[(4-methoxyphenyl)methyl]amino}-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol

The title compound was obtained in the same manner as in Step 1 ofExample 2, using the compound obtained in Step 2 of Reference ExampleC-23 and 4-(H-imidazol-1-yl)benzaldehyde (CAS: 10040-98-9).

¹H-NMR (CDCl₃) δ: 1.75 (1H, dt, J=14.1, 2.5 Hz), 1.88-2.04 (2H, m), 2.23(1H, ddd, J=14.1, 8.6, 6.7 Hz), 3.28-3.34 (1H, m), 3.44 (3H, s), 3.52(2H, q, J=10.2 Hz), 3.78 (3H, s), 3.86 (2H, d, J=1.8 Hz), 4.34 (1H, brs), 4.52 (2H, d, J=4.9 Hz), 4.79 (1H, td, J=9.7, 4.7 Hz), 5.09 (1H, brs), 6.84 (2H, d, J 30=8.6 Hz), 7.20 (2H, d, J=8.6 Hz), 7.24 (2H, d,J=8.6 Hz), 7.27-7.28 (1H, m), 7.36 (2H, d, J=8.6 Hz), 7.43 (2H, d, J=8.6Hz), 7.84 (1H, s).

MS (m/z): 638 (M+H)⁺.

Step 2(1R,2S,4R)-2-{[2-amino-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)cyclopentan-1-ol

To a solution of the compound (137 mg) obtained in the above Step 1 indichloromethane (0.55 mL) was added trifluoroacetic acid (CAS: 76-05-1)(1.64 mL) at room temperature. The mixture was stirred 60° C. for 1.5hr, and the reaction solution was concentrated under reduced pressure.The obtained residue was subjected to silica gel column chromatography(ethyl acetate/methanol) to give the title compound (60.1 mg) as asolid.

¹H-NMR (CD₃OD) δ: 1.59 (1H, ddd, J=14.1, 6.7, 3.1 Hz), 2.10 (1H, td,J=12.3, 8.6 Hz), 2.18-2.25 (1H, m), 2.37-2.45 (1H, m), 3.09-3.21 (1H,m), 3.45 (3H, s), 3.71 (2H, q, J=10.6 Hz), 3.88 (2H, d, J=3.1 Hz),4.47-4.51 (1H, m), 4.81-4.88 (1H, m), 7.15 (1H, s), 7.40 (1H, s), 7.55(4H, s), 7.57-7.58 (1H, m), 8.14 (1H, s).

Step 3(1R,2S,4R)-2-{[2-amino-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)cyclopentan-1-olhydrochloride

The compound (0.601 g) obtained in the above Step 2 was dissolved inethanol (1.5 mL), and 1N hydrochloric acid (0.116 mL) was added thereto.The solvent was evaporated under reduced pressure, and the residue wasdried. The obtained residue was suspended in diethyl ether (3 mL), andthe resulting solid was collected by filtration, and dried to give thetitle compound (0.0605 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.78-1.85 (1H, m), 2.21-2.29 (1H, m), 2.36-2.46 (2H,m), 3.33 (3H, s), 3.51 (1H, br s), 3.89 (2H, q, J=11.0 Hz), 4.24 (2H, brs), 4.35 (1H, br s), 4.74-4.81 (1H, m), 5.17 (1H, d, J=4.3 Hz), 6.16(2H, s), 7.15 (1H, s), 7.43 (1H, s), 7.73 (2H, d, J=8.8 Hz), 7.77 (2H,d, J=8.8 Hz), 7.83 (1H, t, J=1.5 Hz), 8.37 (1H, s), 9.36-9.48 (2H, m).

MS (m/z): 518 (M+H)⁺.

Example 225-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-3-methoxypyridine-2-carbonitrilehydrochloride Step 15-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-3-methoxypyridine-2-carbonitrile

A mixture of the compound (0.0756 g) obtained in Step 2 of ReferenceExample D-25, the compound (0.110 g) obtained in Step 2 of ReferenceExample C-14, sodium triacetoxyborohydride (0.208 g), acetic acid (0.092mL) and dichloromethane (5 mL) was stirred at room temperature for 15hr. The reaction solution was diluted with dichloromethane, washedsuccessively with saturated aqueous sodium bicarbonate solution andsaturated brine, dried over anhydrous magnesium sulfate, andconcentrated under reduced pressure. The obtained residue was subjectedto silica gel column chromatography (ethyl acetate/methanol) to give thetitle compound (0.0707 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.84-1.90 (1H, m), 1.95-2.03 (1H, m), 2.06-2.13 (1H,m), 2.32-2.40 (1H, m), 3.41-3.46 (1H, m), 3.55 (3H, s), 3.63 (2H, q,J=10.2 Hz), 3.89-3.96 (2H, m), 4.05 (3H, s), 4.49-4.53 (1H, m), 5.07(1H, td, J=9.7, 4.5 Hz), 7.40 (1H, s), 7.46 (1H, d, J=1.8 Hz), 7.49 (2H,d, J=8.0 Hz), 7.59 (2H, d, J=8.0 Hz), 8.40 (1H, s), 8.50 (1H, d, J=1.8Hz).

Step 25-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-3-methoxypyridine-2-carbonitrilehydrochloride

A mixture of the compound (0.0707 g) obtained in the above Step 1, 1Nhydrochloric acid (0.120 mL) and ethanol (5.00 mL) was stirred at roomtemperature for 10 min, and concentrated under reduced pressure. Theobtained residue was suspended in acetonitrile (10 mL), and theresulting solid was collected by filtration, and dried to give the titlecompound (0.0683 g) as a solid.

¹H-NMR (DMSO-D₆, 80° C.) δ: 1.82-1.90 (1H, m), 2.26-2.34 (1H, m),2.40-2.51 (2H, m), 3.45 (3H, s), 3.58-3.65 (1H, m), 4.03 (2H, q, J=11.0Hz), 4.09 (3H, s), 4.24-4.31 (2H, m), 4.36-4.40 (1H, m), 4.91-4.98 (1H,m), 5.01-5.09 (1H, m), 7.68 (1H, s), 7.74 (2H, d, J=8.0 Hz), 7.95 (2H,d, J=8.0 Hz), 7.98 (1H, s), 8.34 (1H, s), 8.65-8.66 (1H, m), 9.22-9.38(1H, m).

MS (m/z): 569 (M+H)⁺.

[α]_(D) ²⁰ −22.9 (c 1.00, DMSO)

Example 23(1R,2S,4R)-4-[({4-[1-(methanesulfonyl)-1H-indazol-4-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride Step 1(1R,2S,4R)-4-[({4-[1-(methanesulfonyl)-1H-indazol-4-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

A mixture of the compound (0.0817 g) obtained in Step 2 of ReferenceExample C-14, the compound (0.0708 g) obtained in Reference ExampleD-40, dichloromethane (2.5 mL), DIPEA (0.0411 mL), acetic acid (0.0810mL) and sodium triacetoxyborohydride (0.150 g) was stirred at roomtemperature for 2 hr. Saturated aqueous sodium hydrogencarbonatesolution was added to the reaction mixture, and the mixture wasextracted with chloroform. The organic layer was washed with saturatedbrine, dried over anhydrous sodium sulfate, filtered, and concentratedunder reduced pressure. The obtained residue was subjected to silica gelcolumn chromatography (ethyl acetate/methanol) using amino silica gelcolumn as a charge column to give the title compound (0.105 g) as asolid.

¹H-NMR (CDCl₃) δ: 1.90 (1H, d, J=13.5 Hz), 1.95-2.04 (1H, m), 2.10 (1H,ddd, J=13.5, 6.1, 4.3 Hz), 2.38 (1H, ddd, J=12.9, 9.8, 6.7 Hz), 3.29(3H, s), 3.46-3.54 (1H, m), 3.56 (3H, s), 3.64 (2H, q, J=10.0 Hz),3.90-3.98 (2H, m), 4.49-4.53 (1H, m), 5.12 (1H, td, J=9.8, 4.3 Hz), 7.41(1H, s), 7.43 (1H, d, J=7.4 Hz), 7.50 (2H, d, J=8.0 Hz), 7.61 (2H, d,J=8.0 Hz), 7.64 (1H, dd, J=8.6, 7.4 Hz), 8.09 (1H, d, J=8.6 Hz), 8.40(1H, s), 8.40 (1H, s).

Step 2(1R,2S,4R)-4-[({4-[1-(methanesulfonyl)-1H-indazol-4-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

The compound (0.105 g) obtained in the above Step 1 was dissolved inethanol (1.5 mL), and 1N hydrochloric acid (0.166 mL) was added thereto.The solvent was evaporated under reduced pressure, and the residue wasdried. The obtained residue was suspended in diethyl ether (3 mL), andthe resulting solid was collected by filtration, and dried to give thetitle compound (0.0966 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.81-1.89 (1H, m), 2.28-2.47 (3H, m), 3.45 (3H, s),3.53 (3H, s), 3.63 (1H, br s), 4.10 (2H, q, J=10.4 Hz), 4.27-4.33 (2H,m), 4.35-4.38 (1H, br m), 4.92-5.02 (1H, m), 5.20 (1H, br s), 7.58 (1H,d, J=6.7 Hz), 7.73-7.86 (6H, m), 8.03 (1H, d, J=8.6 Hz), 8.37 (1H, s),8.64 (1H, s), 9.45-9.57 (2H, m).

MS (m/z):631 (M+H)⁺.

[α]_(D) ²⁰ −20.4 (c 1.00, MeOH)

Example 24 N⁴-[(1S,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-methoxycyclopentyl]-N²,N⁴-dimethyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine-2,4-diaminehydrochloride Step 1(1R,3S,4S)—N³-[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]-N¹-{[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}-4-methoxy-N³-methylcyclopentane-1,3-diamine

A mixture of the compound (0.440 g) obtained in Step 2 of ReferenceExample E-2, the compound (0.318 g) obtained in Step 2 of ReferenceExample B-4, 2-propanol (22 mL) and DIPEA (CAS: 7087-68-5) (3.2 mL) wasstirred at 100° C. for 3 hr. Saturated brine/water (3:1) was added tothe reaction solution, and the mixture was extracted with ethyl acetate.The organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (ethyl acetate/methanol) to give the titlecompound (0.508 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.56-1.65 (1H, m), 1.90-2.05 (2H, m), 2.34-2.45 (1H,m), 3.32 (3H, s), 3.35-3.44 (1H, m), 3.36 (3H, s), 3.61 (2H, q, J=10.2Hz), 3.83 (2H, s), 3.95 (3H, s), 4.07 (3H, s), 4.11-4.19 (1H, m),5.01-5.13 (1H, m), 7.25 (1H, d, J=1.8 Hz), 7.38-7.44 (2H, m), 7.47 (1H,s), 7.49-7.55 (2H, m), 7.95 (1H, d, J=1.8 Hz).

MS (m/z): 622, 624 (M+H)⁺.

Step 2N⁴-[(1S,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-methoxycyclopentyl]-N²,N⁴-dimethyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine-2,4-diamine

The compound (0.253 g) obtained in the above Step 1 was dissolved inbutyronitrile (2.5 mL), and methylamine hydrochloride (CAS: 593-51-1)(0.109 g) and DIPEA (CAS: 7087-68-5) (0.42 mL) were added thereto, andthe mixture was subjected to microwave irradiation at 150° C. for 1 hr.Then, additional methylamine hydrochloride (CAS: 593-51-1) (0.106 g) andDIPEA (CAS: 7087-68-5) (0.40 mL) were added thereto, and the mixture wassubjected to microwave irradiation at 160° C. for 3 hr. Water was addedto the reaction solution, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated brine, dried overanhydrous sodium sulfate, and concentrated under reduced pressure, andthe residue was subjected to silica gel column chromatography(dichloromethane/methanol), reverse-phase HPLC (GILSON, water (0.10%formic acid)/acetonitrile (0.10% formic acid)) and amino silica gelcolumn chromatography (ethyl acetate/methanol). The obtained oil wasdissolved in diethyl ether, and the solution was washed with water andsaturated brine, dried over anhydrous sodium sulfate, and concentratedunder reduced pressure to give the title compound (0.0773 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.53-1.58 (1H, m), 1.83-1.93 (1H, m), 1.97-2.06 (1H,m), 2.30-2.39 (1H, m), 2.99 (3H, d, J=4.9 Hz), 3.26 (3H, s), 3.30-3.40(1H, m), 3.31 (3H, s), 3.52 (2H, q, J=10.2 Hz), 3.81 (1H, d, J=13.2 Hz),3.85 (1H, d, J=13.2 Hz), 3.95 (3H, s), 4.06-4.14 (1H, m), 4.07 (3H, s),4.75 (1H, q, J=4.9 Hz), 5.11-5.21 (1H, m), 7.24 (1H, d, J=2.5 Hz), 7.26(1H, s), 7.40 (2H, d, J=8.6 Hz), 7.51 (2H, d, J=8.6 Hz), 7.95 (1H, d,J=2.5 Hz).

MS (m/z): 617(M+H)⁺.

Step 3N⁴-[(1S,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-methoxycyclopentyl]-N²,N⁴-dimethyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine-2,4-diaminehydrochloride

The title compound was obtained in the same manner as in Step 3 ofExample 2, using the compound obtained in the above Step 2.

¹H-NMR (DMSO-D₆) δ: 1.99-2.13 (2H, m), 2.26-2.45 (2H, m), 2.78 (3H, d,J=4.9 Hz), 3.22 (6H, s), 3.58-3.71 (1H, m), 3.82-3.96 (2H, m), 3.90 (3H,s), 3.91 (3H, s), 4.12-4.29 (3H, m), 4.93-5.15 (1H, m), 6.71 (1H, q,J=4.9 Hz), 7.46 (1H, s), 7.58 (1H, d, J=1.8 Hz), 7.67 (2H, d, J=8.0 Hz),7.81 (2H, d, J=8.0 Hz), 8.06 (1H, d, J=1.8 Hz), 9.50 (1H, br s), 9.56(1H, br s).

MS (m/z): 617 (M+H)⁺.

Example 25(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride Step 1(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

A mixture of the compound (0.102 g) obtained in Step 2 of ReferenceExample C-14, the compound (0.0844 g) obtained in Reference ExampleD-26, dichloromethane (3.2 mL), sodium triacetoxyborohydride (CAS:56553-60-7) (0.215 g) and acetic acid (0.0500 mL) was stirred at roomtemperature for 17 hr. Water/saturated aqueous sodium bicarbonatesolution (2/1) was added to the reaction solution, and the mixture wasextracted with dichloromethane/methanol (9/1). The organic layer wasdried over anhydrous sodium sulfate, filtered, and concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (dichloromethane/methanol) to give the title compound(0.115 g) as an oil.

¹H-NMR (CDCl₃) δ: 1.80-1.86 (1H, m), 1.94-2.03 (1H, m), 2.10 (1H, dt,J=13.9, 5.7 Hz), 2.28-2.38 (1H, m), 3.38-3.45 (1H, m), 3.53 (3H, s),3.61 (2H, q, J=10.2 Hz), 3.89 (1H, d, J=13.2 Hz), 3.93 (1H, d, J=13.2Hz), 4.00 (3H, s), 4.23 (3H, s), 4.46-4.51 (1H, m), 5.04 (1H, td, J=9.7,4.7 Hz), 7.11 (1H, s), 7.38 (1H, s), 7.44 (2H, d, J=8.0 Hz), 7.96 (2H,d, J=8.0 Hz), 8.40 (1H, s).

MS (m/z):575 (M+H)⁺.

Step 2(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

The compound (0.112 g) obtained in the above Step 1 was dissolved inethanol (3.6 mL), 1N hydrochloric acid (0.173 mL) was added thereto, andthe solvent was evaporated under reduced pressure. The obtained solidwas suspended in diethyl ether, and the resulting solid was collected byfiltration, and dried to give the title compound (0.104 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.80-1.89 (1H, m), 2.26-2.36 (1H, m), 2.43-2.54 (2H,m), 3.44 (3H, s), 3.55-3.66 (1H, m), 4.01 (3H, s), 4.03-4.16 (5H, m),4.22-4.40 (3H, m), 4.90-5.00 (1H, m), 5.20 (1H, d, J=4.3 Hz), 7.67 (1H,s), 7.71-7.79 (3H, m), 8.19 (2H, d, J=8.0 Hz), 8.36 (1H, s), 9.47 (1H,br s), 9.53 (1H, br s).

MS (m/z):575 (M+H)⁺.

[α]_(D) ²⁰ −23.5 (c 1.00, MeOH)

Example 26(1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride Step 1(1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

A mixture of the compound (0.112 g) obtained in Step 3 of ReferenceExample C-14, dichloromethane (3.0 mL), the compound (0.080 g) obtainedin Step 2 of Reference Example D-11, DIPEA (0.153 mL), acetic acid(0.084 mL) and sodium triacetoxyborohydride (0.190 g) was stirred atroom temperature for 16 hr. Saturated aqueous sodium hydrogencarbonatesolution was added to the reaction mixture, and the mixture wasextracted with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure. The obtained residue was subjectedto silica gel column chromatography (ethyl acetate-methanol) using aminosilica gel column as a charge column to give the title compound (0.125g) as a solid.

¹H-NMR (CDCl₃) δ: 1.46-1.82 (2H, m), 1.86-2.09 (3H, m), 2.30-2.40 (1H,m), 3.03 (6H, s), 3.43-3.50 (1H, m), 3.55 (3H, s), 3.59-3.68 (2H, m),3.83-3.92 (2H, m), 3.93 (3H, s), 4.46-4.51 (1H, m), 5.09-5.18 (1H, m),7.20 (1H, d, J=1.8 Hz), 7.37-7.42 (3H, m), 7.50-7.55 (2H, m), 8.10 (1H,d, J=1.8 Hz), 8.40 (1H, s).

MS (m/z): 587 (M+H)⁺.

Step 2(1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

The compound (0.122 g) obtained in the above Step 1 was dissolved inethanol (2 mL), and 5N aqueous hydrochloric acid solution (0.042 mL) wasadded thereto. The solvent was evaporated under reduced pressure, andthe residue was dried. The obtained residue was suspended inacetonitrile (2 mL), and the resulting solid was collected byfiltration, and dried to give the title compound (0.117 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.79-1.88 (1H, m), 2.25-2.35 (1H, m), 2.38-2.55 (2H,m), 2.97 (6H, s), 3.44 (3H, s), 3.53-3.65 (1H, m), 3.91 (3H, s),4.03-4.16 (2H, m), 4.17-4.30 (2H, m), 4.31-4.39 (1H, m), 4.88-5.01 (1H,m), 5.11-5.30 (1H, m), 7.47-7.51 (1H, m), 7.63-7.69 (2H, m), 7.73-7.81(3H, m), 8.10-8.14 (1H, m), 8.36 (1H, s), 9.31-9.53 (2H, m).

MS (m/z): 587 (M+H)⁺.

[α]_(D) ²⁰ −24.1 (c 1.00, MeOH)

Example 27(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride Step 1(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol

A mixture of the compound (90.0 mg) obtained in Step 2 of ReferenceExample C-15, the compound (55.9 mg) obtained in Reference Example D-26,DIPEA (0.114 mL), dichloromethane (2.18 mL), sodiumtriacetoxyborohydride (139 mg) and acetic acid (0.0624 mL) was stirredat room temperature for 1.5 hr. Saturated aqueous sodiumhydrogencarbonate solution and dichloromethane were added to thereaction solution, and the mixture was subjected to liquid separation.The organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (ethyl acetate/methanol) to give thetitle compound (94.0 mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.49-1.51 (1H, m), 2.01-2.18 (3H, m), 3.07-3.10 (1H,m), 3.38 (3H, s), 3.81-3.83 (5H, m), 3.94-3.97 (5H, m), 4.06 (3H, s),4.29-4.31 (1H, m), 4.72-4.74 (2H, m), 7.46-7.52 (4H, m), 8.00-8.02 (2H,m).

MS (m/z): 605 (M+H)⁺.

Step 2(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

A mixture of the compound (90.0 mg) obtained in the above Step 1,ethanol (1 mL) and 1N hydrochloric acid ethanol solution (0.149 mL) wasstirred at room temperature, and concentrated under reduced pressure.Ethanol/diethyl ether was added to the residue, and the resulting solidwas collected by filtration, and dried to give the title compound (84.1mg) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.85-1.89 (1H, m), 2.29-2.35 (1H, m), 2.42-2.51 (2H,m), 3.42 (3H, s), 3.61-3.66 (1H, m), 3.85 (3H, s), 3.93-4.01 (5H, m),4.08 (3H, s), 4.26-4.28 (2H, m), 4.36-4.39 (1H, m), 4.82-4.88 (1H, m),7.59 (1H, s), 7.62 (1H, s), 7.73 (2H, d, J=8.3 Hz), 8.17 (2H, d, J=8.3Hz), 9.40-9.46 (2H, m).

MS (m/z):605 (M+H)⁺.

[α]_(D) ²⁰ −35.6 (c 1.00, MeOH)

Each product in the following Table 2-1 to Table 2-54 was produced inthe same manner as in Step 1 of Example 27, using Raw Material 1 and RawMaterial 2 described in the tables. When the final form ishydrochloride, it was produced in the same manner as in Step 2 ofExample 27 using the free product as a raw material. In Table 2-1 toTable 2-54, when the final form is shown by two items, the compound ofExample is the salt compound shown in the lower item (for example, thecompound of Example 54 in Table 2-14 is(1S,2S,4R)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}-4-[({4-[(pyridin-3-yl)amino]phenyl}methyl)amino]cyclopentan-1-olhydrochloride, which is shown in the lower item). In ExperimentalExample, the above salt compound was used as the compound of Example.

TABLE 2-11 Fimal Form (Compound Name, NMR, MS) Ex. Product Raw Material1 Raw Material 2 Ex. 28

  CAS: 2043-61-0

  Reference Example C-4 Step 2(1R,3S)-N³-(cyclohexylmethyl)-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 0.92-1.27 (5H, m), 1.60-2.10 (11H, m), 2.24-2.34(1H, m), 2.74-2.79 (2H, m), 3.27 (3H, s), 3.54-3.61 (1H, m), 4.06 (1H,d, J = 12.0 Hz), 4.12 (1H, d, J = 12.0 Hz), 5.21-5.31 (1H, m), 7.72 (1H,s), 8.38 (1H, s), 8.76 (1H, br s), 8.88 (1H, br s). MS (m/z): 427 (M +H)⁺. Ex. 29

   

  CAS: 122-85-0

  Reference Example C-1 Step 2N-[4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclohexyl]amino}methyl)phenyl]acetamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.31-1.44 (3H, m), 1.57 (1H, q, J = 11.9 Hz),1.85-1.95 (2H, m), 2.05 (3H, s), 2.14-2.19 (1H, m), 2.42-2.48 (1H, m),3.13-3.22 (1H, m), 4.06-4.22 (5H, m), 7.48 (2H, d, J = 8.5 Hz), 7.63(2H, d, J = 8.5 Hz), 7.75 (1H, s), 8.44 (1H, s), 8.45 (1H, br s), 9.18(2H, br s), 10.13 (1H, s). MS (m/z): 478 (M + H)⁺.

TABLE 2-2 Ex. 30

  CAS: 82625-45-4

  Reference Example C-4 Step 2(1R,3S)-N¹-methyl-N³-({4-[2-(morpholin-4-yl)ethoxy]phenyl}methyl)-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

¹H-NMR (CD₃OD) δ: 1.25-1.35 (2H, m), 1.91-2.17 (4H, m), 2.21-2.31 (1H,m), 2.48-2.56 (1H, m), 2.71 (3H, br s), 2.94 (1H, br s), 3.30 (3H, s),3.68-3.80 (6H, m), 3.87 (1H, d, J = 10.5 Hz), 3.92 (1H, d, J = 10.5 Hz),4.18-4.24 (4H, m), 5.24-5.34 (1H, m), 7.06 (2H, d, J = 8.5 Hz), 7.46(2H, d, J = 8.5 Hz), 7.63 (1H, s), 8.25 (1H, s). MS (m/z): 550 (M + H)⁺.Ex. 31

  CAS: 1004779-24-1

  Reference Example C-4 Step 22-[2-fluoro-4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenoxy]ethan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.86-2.08 (5H, m), 2.30-2.36 (1H, m), 3.28 (3H, s),3.54-3.60 (1H, m), 3.74 (2H, t, J = 4.9 Hz), 4.04-4.14 (6H, m),5.21-5.30 (1H, m), 7.25 (1H, t, J = 8.5 Hz), 7.33 (1H, d, J = 8.5 Hz),7.51 (1H, dd, J = 12.1, 1.8 Hz), 7.72 (1H, s), 8.37 (1H, s), 9.30 (1H,br s), 9.43 (1H, br s). MS (m/z): 499 (M + H)⁺.

TABLE 2-3 Ex. 32

  CAS: 54605-72-0

  Reference Example C-4 Step 2(1R,3S)-N¹-methyl-N³-[(1-phenyl-1H-pyrazol-4-yl)methyl]-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

¹H-NMR (DMSO-D₆) 1.83-2.18 (4H, m), 2.31-2.45 (2H, m), 3.28 (3H, s),3.61-3.70 (1H, m), 4.02- 4.19 (4H, m), 5.24-5.35 (1H, m), 7.36 (1H, J =7.3 Hz), 7.54 (2H, d, J = 7.9 Hz), 7.72 (1H, s), 7.81 (2H, d, J = 7.3Hz), 7.91 (1H, s), 8.37 (1H, s), 8.63 (1H, s), 9.14 (2H, brs). MS (m/z):487 (M + H)⁺. Ex. 33

  diastereomeric mixture

  CAS: 98-86-2

  Reference Example C-4 Step 2(1R,3S)-N¹-methyl-N³-(1-phenylethyl)-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine (diastereomer mixture)

diastereomeric mixture ¹H-NMR (CDCl₃) δ: 1.38 (1.5H, d, J = 1.8 Hz),1.39 (1.5H, d, J = 1.8 Hz), 1.41-1.64 (2.0H, m), 1.77-2.26 (4.0H, m),2.96-3.06 (1.0H, m), 3.24 (1.5H, s), 3.29 (1.5H, s), 3.61 (2.0H, q, J =10.3 Hz), 3.80-3.87 (1.0H, m), 5.13-5.23 (1.0H, m), 7.23-7.37 (6.0H, m),8.37 (0.5H, s), 8.39 (0.5H, s). MS (m/z): 435 (M + H)⁺.

TABLE 2-4 Ex. 34

  Reference Example C-4 Step 2 Reference Example D-85 Step 2N-{-3-[(ethenesulfonyl)amino]propyl}-4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)benzamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.72 (2H, quint, J = 7.1 Hz), 1.84-1.92 (1H, m),1.99-2.12 (4H, m), 2.31- 2.38 (1H, m), 2.86-2.91 (2H, m), 3.27-3.32 (2H,m), 3.29 (3H, s), 3.57-3.65 (1H, m), 4.09 (2H, q, J = 10.9 Hz),4.19-4.24 (2H, m), 5.22-5.31 (1H, m), 5.97 (1H, d, J = 10.0 Hz), 6.02(1H, d, J = 16.4 Hz), 6.70 (1H, dd, J = 16.4, 10.0 Hz), 7.33 (1H, t, J =6.1 Hz), 7.69 (2H, d, J = 8.5 Hz), 7.73 (1H, s), 7.91 (2H, d, J = 8.5Hz), 8.37 (1H, s), 8.59 (1H, t, J = 5.8 Hz), 9.53 (1H, br s), 9.68 (1H,br s). MS (m/z): 611 (M + H)⁺. Ex. 35

  Reference Example D-23

  Reference Example C-2 Step 2(1S,3R)-N¹-{[4-(1,3-oxazol-2-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.34-1.45 (3H, m), 1.61-1.64 (1H, m), 1.89-1.92 (2H,m), 2.18-2.21 (1H, m), 2.45-2.48 (1H, m), 3.22-3.24 (1H, m), 4.05-4.25(5H, m), 7.42 (1H, s), 7.72 (1H, s), 7.77 (2H, d, J = 8.2 Hz), 8.05 (2H,d, J = 8.2 Hz), 8.21-8.27 (2H, m), 8.38 (1H, s), 9.47-9.53 (2H, m). MS(m/z): 488 (M + H)⁺.

TABLE 2-5 Ex. 36

  CAS: 1003-32-3

  Reference Example C-4 Step 2(1R,3S)-N¹-methyl-N³-[(1,3-thiazol-5-yl)methyl]-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-2.12 (5H, m), 2.30-2.37 (1H, m), 3.28 (3H, s),3.59-3.66 (1H, m), 4.09 (2H, q, J = 11.1 Hz), 4.51 (2H, t, J = 5.5 Hz),5.22-5.31 (1H, m), 7.73 (1H, s), 8.11 (1H, s), 8.38 (1H, s), 9.19 (1H,s), 9.50-9.58 (1H, m), 9.66-9.73 (1H, m). MS (m/z): 428 (M + H)⁺. Ex. 37

  Reference Example D-91 Step 2

  Reference Example C-2 Step 2(1S,3R)-N¹-{[4-(4H-1,2,4-triazol-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.40-1.42 (3H, m), 1.67-1.70 (1H, m), 1.90-1.92 (2H,m), 2.18-2.20 (1H, m), 2.47-2.50 (1H, m), 3.22-3.25 (1H, m), 4.10-4.23(5H, m), 7.72 (2H, d, J = 8.3 Hz), 7.84 (1H, s), 8.08 (2H, d, J = 8.3Hz), 8.53-8.55 (2H, m), 8.86 (1H, s), 9.49-9.51 (2H, m). MS (m/z): 488(M + H)⁺.

TABLE 2-6 Ex. 38

  Reference Example D-86 Step 3

  Reference Example C-4 Step 21-{[4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl[amino}cyclohexy]amino}methyl)phenyl]methyl}pyrrolidin-2-onehydrochloride

¹H-NMR (DMSO-D₆) δ: 1.24-1.58 (4H, m), 1.88-1.96 (4H, m), 2.14-2.19 (1H,m), 2.29 (2H, t, J = 8.3 Hz), 2.41-2.49 (1H, m), 3.16-3.25 (3H, m), 4.08(2H, q, J = 11.2 Hz), 4.13-4.21 (3H, m), 4.38 (2H, s), 7.28 (2H, d, J =8.0 Hz), 7.56 (2H, d, J = 8.0 Hz), 7.69 (1H, d, J = 3.7 Hz), 8.10 (1H,br s), 9.23 (2H, br s). MS (m/z): 518 (M + H)⁺. Ex. 39

  racemate Reference Example D-87 Step 3

  Reference Example C-2 Step 2 diastereomeric mixture5-[4-({[(1S,3R)-3-{[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl[amino}cyclohexyl]amino}methyl)phenyl]pyrrolidin-2-one hydrochloride(diastereomer mixture)

diastereomeric mixture ¹H-NMR (DMSO-D₆) δ: 1.25-1.62 (4H, m), 1.66-1.76(1H, m), 1.85-1.95 (2H, m), 2.14-2.20 (1H, m), 2.23 (2H, t, J = 8.0 Hz),2.42-2.48 (2H, m), 3.20 (1H, br s), 4.08 (2H, q, J = 11.2 Hz), 4.10-4.26 (3H, m), 4.69 (1H, t, J = 7.1 Hz), 7.36 (2H, d, J = 8.0 Hz),7.53-7.57 (2H, m), 7.68-7.70 (1H, m), 8.01-8.22 (2H, m), 8.32-8.38 (1H,m), 9.00-9.32 (2H, m). MS (m/z): 504 (M + H)⁺.

TABLE 2-7 Ex. 40

  CAS: 99163-12-9

  Reference Example C-2 Step 2(1S,3R)-N¹-{[4-(pyridin-4-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆, 50° C.) δ: 1.34-1.45 (3H, m), 1.58 (1H, br s),1.89-1.96 (2H, br m), 2.04- 2.08 (1H, m), 2.18-2.21 (1H, br m), 3.25(1H, br s), 4.03 (2H, q, J = 10.9 Hz), 4.21 (1H, br s), 4.26 (2H, br s),7.68 (1H, s), 7.76 (2H, d, J = 6.1 Hz), 7.89 (2H, s), 7.93 (2H, d, J =7.9 Hz), 7.97 (1H, br s), 8.35 (1H, s), 8.73 (2H, d, J = 4.9 Hz), 9.28(2H, br s). MS (m/z): 498 (M + H)⁺. Ex. 41

  CAS: 67-36-7

  Reference Example C-2 Step 2(1S,3R)-N¹-[(4-phenoxyphenyl)methyl]-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.29-1.66 (4H, m), 1.85-1.97 (3H, m), 2.17 (1H, brs), 3.22 (1H, br s), 4.02-4.25 (5H, m), 7.02 (2H, d, J = 8.0 Hz), 7.06(2H, d, J = 9.2 Hz), 7.18 (1H, t, J = 7.4 Hz), 7.42 (2H, t, J = 8.0 Hz),7.56-7.63 (2H, m), 7.72 (1H, d, J = 15.3 Hz), 8.40 (1H, d, J = 11.7 Hz),9.05-9.37 (2H, m). MS (m/z): 513 (M + H)⁺.

TABLE 2-8 Ex. 42

  CAS: 127406-55-7

Reference Example C-9 Step 22-{[(1R,3S)-3-({[4-(pyridin-3-yl)phenyl]methyl}amino)cyclopentyl][6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl[amino}ethan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.92-1.94 (1H, m), 2.07-2.09 (4H, m), 2.39-2.40 (1H,m), 3.69-3.74 (6H, m), 4.11-4.14 (2H, m), 4.23-4.25 (2H, m), 5.03-5.05(1H, m), 7.64 (1H, s), 7.70-7.71 (1H, m), 7.76 (2H, d, J = 8.2 Hz), 7.89(2H, d, J = 8.2 Hz), 8.36-8.38 (2H, m), 8.69-8.71 (1H, m), 9.06 (1H, s),9.54-9.60 (2H, m). MS (m/z): 528 (M + H)⁺. Ex. 43

  CAS: 127406-55-7

  Reference Example C-5 Step 2(1R,3S)-N¹-ethyl-N³-{[4-(pyridin-3-yl)phenyl]methyl}-N¹-[6-(2,2,2trifluoroethyl)thieno[2,3- d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

¹H-NMR (DMSO-D₆) δ: 1.29 (3H, t, J = 6.7 Hz), 1.91-2.07 (5H, m),2.36-2.43 (1H, m), 3.61-3.70 (1H, m), 3.71-3.82 (2H, m), 4.12 (2H, q, J= 10.9 Hz), 4.24 (2H, br s), 5.07-5.16 (1H, m), 7.53- 7.58 (2H, m), 7.74(2H, d, J = 7.9 Hz), 7.85 (2H, d, J = 7.9 Hz), 8.16 (1H, d, J = 7.9 Hz),8.37 (1H, s), 8.62 (1H, d, J = 3.0 Hz), 8.96 (1H, s), 9.47 (1H, br s),9.62 (1H, br s). MS (m/z): 512 (M + H)⁺.

TABLE 2-9 Ex. 44

  Reference Example D-88 Step 3

  Reference Example C-2 Step 2(1S,3R)-N¹-{[4-(cyclohexylamino)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.13-1.21 (2H, m), 1.24-1.52 (6H, m), 1.55-1.63 (1H,m), 1.66-1.76 (3H, m), 1.82-1.96 (5H, m), 2.11-2.18 (1H, m), 2.32-2.44(1H, m), 3.20 (1H, br s), 3.99 (2H, br s), 4.08 (2H, q, J = 11.0 Hz),4.14-4.22 (1H, m), 5.60-5.72 (1H, m), 6.55-6.68 (2H, m), 7.15-7.22 (2H,m), 7.66 (1H, d, J = 4.9 Hz), 7.96-8.08 (1H, m), 8.35 (1H, s), 8.53-8.73(2H, m). MS (m/z): 518 (M + H)⁺. Ex. 45

  Reference Example D-65

  Reference Example C-12 Step 2(1R,3S)-N¹-(6-cyclopropylthieno[2,3-d]pyrimidin-4-yl)-N³-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N¹-methylcyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 0.79-0.81 (2H, m), 1.06-1.09 (2H, m), 1.88-2.45 (7H,m), 3.26 (3H, s), 3.64-3.67 (1H, m), 3.92 (3H, s), 4.21-4.24 (2H, m),5.16-5.19 (1H, m), 7.27 (1H, s), 7.62-7.62 (1H, m), 7.69 (2H, d, J = 8.3Hz), 7.82 (2H, d, J = 8.3 Hz), 8.28 (1H, s), 8.31-8.31 (1H, m), 8.51-8.51 (1H, m), 9.25-9.35 (2H, m). MS (m/z): 486 (M + H)⁺.

TABLE 2-10 Ex. 46

Reference Example D-71

Reference Example C-10 Step 2(1R,3S)-N³-[(5′-methoxy[2,3′-bipyridine]-5-yl)methyl]-N¹-methyl-N¹-[2-methyl-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.86-2.16 (5H, m), 2.31-2.42 (1H, m), 2.46 (3H, s),3.27 (3H, s), 3.68 (1H, br s), 3.94 (3H, s), 4.05 (2H, q, J = 11.2 Hz),4.30 (2H, br s), 5.24-5.35 (1H, m), 7.66 (1H, s), 8.03-8.06 (1H, m),8.16 (1H, dd, J = 8.0, 2.5 Hz), 8.23 (1H, d, J = 8.0 Hz), 8.41 (1H, d, J= 2.5 Hz), 8.88 (1H, s), 8.93 (1H, d, J = 1.8 Hz), 9.36-9.53 (2H, m). MS(m/z): 543 (M + H)⁺. Ex. 47

Reference Example D-65

Reference Example C-8 Step 2(1R,3S)-N¹-[6-(methoxymethyl)thieno[2,3-d]pyrimidin-4-yl]-N³-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N¹-methylcyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.91-1.93 (1H, m), 2.03-2.10 (4H, m), 2.35-2.42 (1H,m) 3.29 (3H, s), 3.34 (3H, s), 3.63-3.65 (1H, m), 3.92 (3H, s), 4.21(2H, s), 4.67 (2H, s), 5.22-5.24 (1H, m), 7.57- 7.70 (6H, m), 8.31-8.32(2H, m), 8.51 (1H, s), 9.49-9.61 (2H, m). MS (m/z): 490 (M + H)⁺.

TABLE 2-11 Ex. 48

Reference Example D-65

Reference Example C-11 Step 2(1R,3S)-N³-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(oxetan-3-yl)thieno[2,3-d]pyrixnidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆ )δ: 1.56-1.62 (2H, m), 1.78-1.92 (3H, m), 2.14-2.20 (2H,m), 3.13-3.17 (2H, m), 3.42-3.48 (1H, m), 3.70-3.82 (5H, m), 3.89-3.93(4H, m), 3.98-4.01 (1H, m), 4.88-4.91 (1H, m), 5.15-5.24 (1H, m), 7.45(1H, s), 7.47 (2H, d, J = 8.2 Hz), 7.56-7.56 (1H, m), 7.66 (2H, d, J =8.2 Hz), 8.27-8.28 (2H, m), 8.46 (1H, s). Ex. 49

Reference Example C-4Step 2 Reference Example D-84N-[(1-methyl-1H-pyrazol-4-yl)methyl]-4-({[(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)benzamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.84-2.15 (5H, m), 2.31-2.39 (1H, m), 3.27 (3H, s),3.63 (1H, br s), 3.78 (3H, s), 4.09 (2H, q, J = 11.0 Hz), 4.23 (2H, t, J= 6.1 Hz), 4.30 (2H, d, J = 5.5 Hz), 5.27 (1H, br s), 7.36 (1H, s), 7.61(1H, s), 7.63 (2H, d, J = 8.0 Hz), 7.72 (1H, s), 7.93 (2H, d, J = 8.0Hz), 8.37 (1H, s), 8.90 (1H, t, J = 5.5 Hz), 9.41-9.55 (2H, m). MS(m/z): 558 (M + H)⁺.

TABLE 2-12 Ex. 50

Reference Example D-65

Reference Example C-30 Step 2(1R,2R,4S)-2-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.80-1.91 (1H, m), 1.95-2.08 (1H, m), 2.25-2.35 (1H,m), 2.37-2.46 (1H, m), 3.27 (3H, s), 3.35-3.40 (1H, m), 3.94 (3H, s),4.09 (2H, q, J = 10.7 Hz), 4.24-4.36 (2H, m), 4.44-4.54 (1H, m),5.44-5.56 (2H, m), 7.69-7.80 (4H, m), 7.85-7.91 (2H, m), 8.31-8.41 (2H,m), 8.53-8.60 (1H, m), 9.52-9.79 (2H, m). MS (m/z): 544 (M + H)⁺. Ex. 51

Reference Example D-65

Reference Example C-31 Step 2(1S,2R,4S)-2-({[4(5methoxypyridin-3-yl)phenyl]methyl}amino)-4-{methyl[6-(2,2,2-trilluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (CD₃OD, 50° C. δ: 1.96-2.03 (1H, m), 2.21-2.31 (1H, m), 2.42-2.55(2H, m), 3.46 (3H, s), 3.56-3.64 (1H, m), 3.87 (2H, q, J = 10.5 Hz),3.97 (3H, s), 4.32 (1H, d, J = 12.8 Hz), 4.39 (1H, d, J = 12.8 Hz),4.46-4.49 (1H, m), 5.43-5.52 (1H, m), 7.61 (1H, s), 7.65 (1H, s), 7.67(2H, d, J = 8.5 Hz), 7.80 (2H, d, J = 8.5 Hz), 8.28 (1H, d, J = 2.4 Hz),8.32 (1H, s), 8.42 (1H, d, J = 2.4 Hz).

TABLE 2-13 Ex. 52

Reference Example D-65

Reference Example C-13 Step 2(1R,3S)-5,5-difluoro-N¹-{[4-(5-methoxypyridin-3-yl)phenyl]methyl}-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-l,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-1.84 (1H, m), 1.91-2.08 (1H, m), 2.17-2.34 (1H,m), 2.57-2.66 (3H, m), 2.78-2.81 (1H, m), 3.94 (3H, s), 4.11 (2H, q, J =11.0 Hz), 4.33 (2H, s), 4.41-4.43 (1H, m), 7.64 (1H, s), 7.72 (2H, d, J= 8.3 Hz), 7.79 (1H, s), 7.89 (2H, d, J = 8.3 Hz), 8.26-8.28 (1H, m),8.38 (1H, s), 8.42 (1H, s), 8.59 (1H, s), 9.66-9.73 (2H, m), MS (m/z):564 (M + H)⁺. Ex. 53

CAS:66-39-7 Reference Example C-4 Step 2(1R,3S)N³-{[4-(benzenesulfonyl)phenyl]methyl}N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.89-1.98 (5H, m), 2.33-2.33 (1H, m), 3.25 (3H, s),3.61-3.63 (1H, m), 4.07-4.10 (2H, m), 4.23-4.25 (2H, m), 5.23-5.25 (1H,m), 7.62-7.66 (2H, m), 7.70-7.71 (2H, m), 7.80 (2H, d, J = 8.0 Hz),7.97-8.00 (2H, m), 8.07 (2H, d, J = 8.0 Hz), 8.36 (1H, s), 9.26-9.39(2H, m). MS (m/z): 561 (M + H)⁺.

TABLE 2-14 Ex. 54

Reference Example D-80

Reference Example C-22 Step 3(1S,2S,4R)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyriimdin-4-yl]amino}-4-[({4-[(pyridin-3yl)amino]phenyl}methyl)amino]cyclopentan-1-ol

¹H-NMR (DMSO-D₆) δ: 1.44-1.48 (1H, m), 1.61-1.68 (1H, m), 1.81-1.84 (1H,m), 2.18-2.25 (1H, m), 2.77 (3H, d, J = 4.9 Hz), 3.05-3.15 (4H, m), 3.60(2H, s), 3.89 (2H, q, J = 11.2 Hz), 4.32-4.35 (1H, m), 4.71-4.74 (1H,m), 4.88-4.89 (1H, m), 6.57-6.58 (1H, m), 7.04-7.06 (2H, m), 7.20 7.24(3H, m), 7.41-7.42 (2H, m), 7.98-8.00 (1H, m), 8.30-8.31 (2H, m). MS(m/z): 558 (M + H)⁺,

¹H-NMR (DMSO-D₆) δ: 1.85-1.96 (2H, m), 2.19-2.36 (2H, m), 2.79 (3H, d, J= 4.9 Hz), 3.19 (3H s), 3.62-3.65 (1H, m), 3.90 (2H, q, J = 11.2 Hz),4.05 (2H, s), 4.48-4.50 (1H, m), 4.72-4.75 (1H, m), 5.20-5.22 (1H, m),6.71 (1H, d, J = 4.3 Hz), 7.15 (2H, d, J = 8.6 Hz), 7.31-7.33 (1H, m),7.46- 7.48 (3H, m), 7.53-7.56 (1H, m), 8.09-8.10 (1H, m), 8.38-8.39 (1H,m), 8.69 (1H, s), 9.28-9.43 (2H, m). MS (m/z): 558 (M + H)⁺.

TABLE 2-15 Ex. 55

Reference Example D-83 Step 4 Reference Example C-7 Step 2N-{3-[4-({[(1S,3R)-3{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4- yl]amino}cyclopentyl]amino}methyl)anilino]phenyl}prop-2-enamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.84-2.09 (5H, m), 2.31-2.39 (1H, m), 2.80 (3H, s),3.19 (3H, s), 3.40-3.48 (1H, m), 3.51-3.59 (1H, m), 3.91 (2H, q, J =11.1 Hz), 4.04-4.09 (2H, m), 5.09-5.17 (1H, m) ,5.73-5.77 (1H, m), 6.23(1H, d, J = 17.0 Hz), 6.42-6.49 (1H, m), 6.76-6.79 (1H, m), 7.02-7.05(1H, m), 7.11-7.21 (3H, m), 7.38-7.45 (3H, m), 7.74 (1H, s), 8.44 (1H,s), 9.16 (2H, br s), 10.13-10.15 (1H, m). MS (m/z): 610 (M + H)⁺. Ex. 56

Reference Example D-81 Step 2 Reference Example C-4 Step 2(1R,3S)-N³-{[4-(2)-3-dihydro-1H-indol-1-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-1.93 (1H, m), 1.95-2.12 (4H, m), 2.30-2.38 (1H,m), 3.11 (2H, t, J = 8.3 Hz), 3.29 (3H, s), 3.61 (1H, br s), 3.95 (2H,t, J = 8.3 Hz), 4.03-4.14 (4H, m), 5.27 (1H, br s), 6.76 (1H, t, J = 7.4Hz), 7.07 (1H, t, J = 7.4 Hz), 7.15 (1H, d, J = 8.0 Hz), 7.20 (1H, d, J= 7.4 Hz), 7.29 (2H, d, J = 8.6 Hz), 7.55 (2H, d, J = 8.6 Hz), 7.73 (1H,s), 8.38 (1H, s), 9.23-9 48 (2H m). MS (m/z): 538 (M + H)⁺.

TABLE 2-16 Ex. 57

Reference Example C-4 Step 2 Reference ExampleD-92l-[4-({[(1S,3R)-3-{methyl[6-(2,2,2trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)benzene-1-sulfonyl]piperidin-3-ol hydrochloride (diastereomer mixture)

¹H-NMR (DMSO-D₆) δ: 1.05-1.11 (1H, m), 1.44-1.49 (1H, m), 1.70-2.15 (8H,m), 2.31-2.36 (2H, m), 3.25-3.39 (4H, m), 3.42-3.44 (1H, m), 3.52-3.56(2H, m), 3.64-3.67 (1H, m), 4.08-4.11 (2H, m), 4.28-4.30 (2H, m),5.26-5.28 (1H, m), 7.73 (1H, s), 7.83-7.85 (4H, m), 8.37 (1H, s),9.54-9.61 (2H, m). MS (m/z): 584 (M + H)⁺. Ex. 58

Reference Example D-34

Reference Example C-4 Step 2(1R,3S)-N³-({4-[5-(difluoromethoxy)-6-methylpyridin-3-yl]phenyl}methyl)-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.88-2.12 (5H, m), 2.33-2.41 (1H, m), 3.29 (3H, s),3.66 (1H, br s), 4.09 (2H, q, J = 11.1 Hz), 4.24 (2H, br s), 5.28 (1H,t, J = 8.8 Hz), 7.41 (1H, t, J = 73.8 Hz), 7.70 7.73 (3H, m), 7.88 (2H,d, J = 8.5 Hz), 7.92 (1H, s), 8.38 (1H, s), 8.73 (1H, d, J = 1.8 Hz),9.25-9.46 (2H, m). MS (m/z): 578 (M + H)⁺.

TABLE 2-17 Ex. 59

Reference Example D-93 Reference Example C-4 Step 2(1R,3S)-N³-{[4-(azepane-1-sulfonyl)phenyl]methyl)-N¹methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.48-1.51 (4H, m), 1.61-1.63 (4H, m), 1.90-2.03 (5H,m), 2.28-2.35 (1H, m), 3.20-3.21 (4H, m), 3.28 (3H, s), 3.62-3.64 (1H,m), 4.08-4.10 (2H, m), 4.24-4.27 (2H, m), 5.26-5.28 (1H, m), 7.72-7.87(5H, m), 8.37 (1H, s), 9.47-9.61 (2H, m). MS (m/z): 582 (M + H)⁺. Ex. 60

Reference Example D-36

Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(5-methoxy-6-methylpyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.59-2.16 (5H, m), 2.31-2.41 (1H, m), 2.51 (3H, s),3.42-3.49 (1H, m), 3.55 (3H, s), 3.58-3.68 (2H, m), 3.87-3.94 (5H, m),4.46-4.52 (1H, m), 5.08-5.17 (1H, m), 7.24-7.25 (1H, m), 7.40-7.44 (3H,m), 7.54-7.59 (2H, m), 8.29-8.31 (1H, m), 8.40 (1H, s). MS (m/z): 558(M + H)⁺.(1R,2S,4R)-4-({[4-(5-methoxy-6-methylpyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.75-1.85 (1H, m), 2.08 (1H, q, J = 11.0 Hz),2.24-2.43 (5H, m), 3.35 (3H, s), 3.41-3.47 (1H, m), 3.93 (3H, s), 4.08(2H, q, J = 11.1 Hz), 4.25 (2H, br s), 4.37 (1H, br s), 5.38-5.50 (1H,m), 6.20 (1H, d, J = 4.0 Hz), 7.60 (1H, s), 7.65-7.77 (3H, m), 7.85 (2H,d, J = 8.0 Hz), 8.33-8.40 (2H, m), 9.02 (1H, br s), 9.38 (1H, br s). MS(m/z): 558 (M + H)⁺.

TABLE 2-18 Ex. 61

CAS:10040-98-9

Reference Example C-20 Step 3(1R,2S,4R)-4-({[4-(1H-imidazol-1-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.77-1.84 (1H, m), 2.25-2.45 (3H, m), 2.77 (3H, d, J= 4.3 Hz), 3.35 (3H, s), 3.49-3.61 (1H, m), 3.89 (2H, q, J = 10.8 Hz),4.20-4.30 (2H, m), 4.36-4.41 (1H, m), 4.73 4.82 (1H, m), 5.17 (1H, d, J= 4.3 Hz), 6.59-6.68 (1H, m), 7.30 (1H, s), 7.43 (1H, s), 7.73-7.82 (4H,m), 7.93 (1H, s), 8.63 (1H, s), 9.40 (2H, br s), MS (m/z): 532 (M + H)⁺.Ex. 62

Reference Example D-75

Reference Example C-20 Step 3(1R,2S,4R)-4[({4[1-(methanesulfonyl)-1H-pyrazol-4-yl]phenyl}methyl)amino]-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.85-1.88 (1H, m), 2.24-2.31 (1H, m), 2.40-2.44 (2H,m), 2.79 (3H, s), 3.35 (3H, s), 3.54-3.57 (4H, m), 3.82-3.85 (2H, m),4.19-4.21 (2H, m), 4.37-4.39 (1H, m), 4.75- 4.81 (1H, m), 5.00-5.02 (1H,m), 6.37 (1H, s), 7.38 (1H, s), 7.63 (2H, d, J = 8.3 Hz), 7.82 (2H, d, J= 8.3 Hz), 8.45 (1H, s), 8.75 (1H, s), 9.36-9.39 (2H, m). MS (m/z): 610(M + H)⁺.

TABLE 2-19 Ex. 63

Reference Example D-42 Reference Example C-4 Step 2(1R,3S)-N³-{[4-(2,3-dihydro[l,4]dioxino[2,3-b]pyridin-7-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.85-1.92 (1H, m), 1.96-2.13 (4H, m), 2.32-2.40 (1H,m), 3.29 (3H, 3.59-3.66 (1H, m), 4.09 (2H, q, J = 11.0 Hz), 4.20 (2H, t,J = 6.7 Hz), 4.29-4.32 (2H, m), 4.44-4.46 (2H, m), 5.23-5.30 (1H, m),7.65 (2H, d, J = 8.0 Hz), 7.66 (1H, s), 7.73 (1H, s), 7.77 (2H J = 8.0Hz), 8.13 (1H, d, J = 1.8 Hz), 8.38 (1H, s), 9.30-9.51 (2H, m). MS(m/z): 556 (M + HH)⁺. Ex. 64

Reference Example D-51

Reference Example C-20 Step 3(1R,2S,4R)-4[({4-[1-(methanesulfonyl)-1H-pyrrolo[2,3-c]pyridin-4yl]phenyl}methyl)amino]-2-{methyl[2-(methylamino)-6-(2,2,2trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino} cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.80-1.90 (1H, m), 2.31 (1H, dd, J = 12.3, 5.5 Hz),2.38-2.49 (2H, m), 2.78 (3H, d, J = 4.3 Hz), 3.37 (3H, s), 3.53-3.64(1H, m), 3.68 (3H, s), 3.90 (2H, q, J = 11.0 Hz), 4.30 (2H, br s), 4.40(1H, br s), 4.80 (1H, br s), 5.17 (1H, br s), 6.62-6.73 (1H, m), 6.97(1H J = 3.7 Hz), 7.45 (1H, s), 7.75-7.82 (4H, m), 7.96 (1H, d, J = 3.7Hz), 8.57 (1H, s), 9.17 (1H, 9.41-9.52 (2H, m). MS (ra/z): 660 (M + H)⁺.

TABLE 2-20 Ex. 65

Reference Example D-39

Reference Example C-14 Step 2(1R,2S,4R)-4({[4(6-chloro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.87 (1H, d, J = 14.1 Hz), 1.97 (1H, ddd, J = 13.5,9.2, 3.1 Hz), 2.03-2.11 (1H, m), 2.36 (1H, ddd, J = 13.5, 9.2, 6.1 Hz),3.41-3.47 (1H, m), 3.55 (3H, s), 3.63 (2H, q, J = 10.0 Hz), 3.90 (2H, d,J = 3.7 Hz), 4.00 (3H, s), 4.48-4.51 (1H, m), 5.10 (1H, td, J = 9.2, 4.9Hz), 7.35 (1H, d, J = 1.8 Hz), 7.40 (1H, s), 7.44 (2H, d, J = 8.0 Hz),7.55 (2H, d, J = 8.0 Hz), 8.20 (1H, d, J = 1.8 Hz), 8.40 (1H, s).(1R,2S,4R)-4({[4-(6-chloro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-1.90 (1H, m), 2.26 2.34 (1H, m), 2.42 2.49 (2H,m), 3.44 (3H, s), 3.59 (1H, br s), 4.02 (3H, s), 4.09 (2H, q, J = 11.0Hz), 4.26 (2H, br s), 4.35 (1H, br s), 4.90-4.99 (1H, m), 5.20 (1H, brs), 7.73-7.78 (3H, m), 7.84 (1H, d, J = 2.5 Hz), 7.91 (2H, d, J = 8.0Hz), 8.35 (1H, d, J = 1.8 Hz), 8.36 (1H, s), 9.45-9.67 (2H, m). MS(m/z): 578 (M + H)⁺. Ex. 66

Reference Example D-66

Reference Example C-4 Step 2N-methyl-4′-({[|(1S,3R)-3-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)[1,1′-biphenyl]-3-carboxamide hydrochloride

1H-NMR (DMSO-D₆) δ: 1.89-1.91 (1H, m), 2.02-2.09 (4H, m), 2.34-2.41 (1H,m), 2.82 (3H, d, J = 4.9 Hz), 3.30 (3H, s), 3.63-3.66 (1H, m), 4.08-4.11(2H, m), 4.21-4.24 (2H, m), 5.27-5.29 (1H, m), 7.57-7.58 (1H, m),7.71-7.73 (3H, m), 7.85-7.86 (4H, m), 8.17 (1H, s), 8.38 (1H, s),8.61-8.62 (1H, m), 9.43-9.56 (2H, m). MS (m/z): 554 (M + H)⁺.

TABLE 2-21 Ex. 67

Reference Example D-56

Reference Example C-14 Step 2(1R,2S,4R)-4-({[4(5,6-dimethoxypyridin-3yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ:1.86-1.99 (2H, m), 2.01-2.08 (1H, m), 2.31-2.40 (1H,m), 3.43-3.48 (1H, m), 3.55 (3H, s), 3.63 (2H, q, J = 10.2 Hz), 3.89(2H, d, J = 4.9 Hz), 3.95 (3H, s), 4.07 (3H, s), 4.47-4.50 (1H, m), 5.13(1H, td, J = 9.8, 4.9 Hz), 7.24 (1H, d, J = 1.8 Hz), 7.40 (2H, d, J =8.0 Hz), 7.41 (1H, s), 7.53 (2H, d, J = 8.0 Hz), 7.95 (1H, d, J = 1.8Hz), 8.40 (1H, s).(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.74-1.86 (1H, m), 2.25-2.47 (3H, m), 3.44 (3H, s),3.57 (1H, br s), 3.90 (3H, s), 3.91 (3H, s), 4.09 (2H, q, J = 11.0 Hz),4.23 (2H, br s), 4.33-4.39 (1H, m), 4.94 (1H, b rs), 5.20 (1H, br s),7.57 (1H, d, J = 1.8 Hz), 7.63-7.70 (2H, m), 7.75 (1H, s), 7.80 (2H, d,J = 8.0 Hz), 8.06 (1H, d, J = 1.8 Hz), 8.36 (1H, s), 9.17-9.26 (2H, m).MS (m/z): 574 (M + H)⁺. Ex. 68

Reference Example D-68

Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(6-fluoro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.78-1.87 (1H, m), 2.25-2.36 (1H, m), 2.37-2.49 (2H,m), 3.44 (3H, s), 3.53-3.64 (1H, m), 4.00 (3H, s), 4.05-4.15 (2H, m),4.22-4.31 (2H, m), 4.32-4.40 (1H, m), 4.88- 4.99 (1H, m), 5.15-5.25 (1H,m), 7.68-7.74 (2H, m), 7.76 (1H, s), 7.85-7.90 (2H, m), 7.90-7.95 (1H,m), 8.07-8.10 (1H, m), 8.36 (1H, s), 9.30-9.50 (2H, m). MS (m/z) : 562(M + H)⁺.

TABLE 2-22 Ex. 69

Reference Example D-77

Reference Example C-20 Step 34′-({[(1R,3R,4S)-3-hydroxy-4-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)-N,N-dimethyl[1,1′-biphenyl]-3-sulfonamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.87-1.90 (1H, m), 2.28-2.46 (3H, m), 2.69 (6H, s),2.82 (3H, s), 3.38 (3H, s), 3.57-3.59 (1H, m), 3.85-3.88 (2H, m),4.25-4.27 (2H, m), 4.40-4.41 (1H, m), 4.77 4.83 (1H, m), 7.43 (1H, s),7.72-8.04 (8H, m), 9.41-9.43 (2H, m). MS (m/z): 649 (M + H)⁺. Ex. 70

Reference Example D-54

Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(4-chloro-1H-pyrrolo[3,2-c]pyridin-7-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-1.88 (1H, m), 2.29-2.37 (1H, m), 2.43-2.49 (2H,m), 3.46 (3H, s), 3.61 (1H, br s), 4.10 (2H, q, J = 11.0 Hz), 4.31 (2H,br s), 4.36 (1H, br s), 4.90-5.01 (1H, m), 6.68 (1H, dd, J = 3.1, 1.8Hz), 7.60 (1H, dd, J = 3.1, 2.5 Hz), 7.74-7.77 (1H, m), 7.77-7.79 (4H,m), 8.05 (1H, s), 8.38 (1H, s), 9.47-9.57 (2H, m), 12.03 (1H, s). MS(m/z): 587, 589 (M + H)⁺.

TABLE 2-23 Ex. 71

Reference Example D-49

Reference Example C-14 Step 2N-[4′-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)[1,1′-biphenyl]-3-yl]-N-methylmethanesulfonamide

¹H-NMR (CDCl3) δ: 1.82-2.11 (3H, m), 2.30-2.41 (1H, m), 2.89 (3H, s),3.39 (3H, s), 3.41-3.49 (1H, m), 3.55 (3H, s), 3.58-3.68 (2H, m),3.84-3.95 (2H, m), 4.44-4.52 (1H, m), 5.08-5.18 (1H, m), 7.33-7.38 (1H,m), 7.38-7.43 (3H, m), 7.44-7.50 (1H, m), 7.50-7.62 (4H, m), 8.40 (1H,s). MS (m/z): 620 (M + H)⁺.N-[4′-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)[1,1′-biphenyl]-3-yl]-N-methylmethanesulfonamidehydrochloride

¹H-NMR (DMSO-D₆) δ: 1.78-1.87 (1H, m), 2.27-2.34 (1H, m), 2.40-2.52 (2H,m), 3.00 (3H, s), 3.31 (3H, s), 3.44 (3H, s), 3.51-3.67 (1H, m),4.02-4.15 (2H, m), 4.20-4.30 (2H, m), 4.33-4.40 (1H, m), 4.87-5.00 (1H,m), 5.08-5.34 (1H, m), 7.42-7.47 (1H, m), 7.50-7.56 (1H, m), 7.63-7.72(4H, m), 7.74-7.82 (3H, m), 8.37 (1H, s), 9.28-9.49 (2H, m). MS (m/z):620 (M + H)⁺.

TABLE 2-24 Ex. 72

Reference Example D-38

Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.76-1.87 (1H, m), 2.29-2.47 (3H, m), 3.45 (3H, s),3.64 (1H, br s), 4.09 (2H, q, J = 11.0 Hz), 4.25 (3H, s), 4.31 (2H, brs), 4.38 (1H, br s), 4.91-5.00 (1H, m), 5.19-5.23 (1H, m), 7.75-7.80(3H, m), 7.93 (2H, d, J = 8.0 Hz), 8.36 (1H, s), 8.37 (1H, s), 8.47 (1H,s), 9.21- 9.41 (2H, m), 9.24 (1H, s). MS (m/z): 568 (M + H)⁺.

TABLE 2-25 Ex. 73

Reference Example D-56

Reference Example C-20 Step 3(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.84 (1H, d, J = 14.1 Hz), 1.95-2.09 (2H, m), 2.31(1H, ddd, J = 13.5, 9.2, 6.7 Hz), 2.96 (3H, d, J = 4.9 Hz), 3.36-3.43(1H, m), 3.46 (3H, s), 3.51 (2H, q, J = 10.4 Hz), 3.88 (2H, d, J = 1.8Hz), 3.95 (3H, s), 4.07 (3H, s), 4.46-4.50 (1H, m), 4.74 (1H, dt, J =4.9, 4.9 Hz), 4.83-4.90 (1H, m), 7.18 (1H, s), 7.24 (1H, d, J = 1.8 Hz),7.41 (2H, d, J = 8.6 Hz), 7.52 (2H, d, J = 8.6 Hz), 7.95 (1H, d, J = 1.8Hz).(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.75-1.86 (1H, m), 2.20-2.46 (3H, m), 2.78 (3H, d, J= 4.3 Hz), 3.36 (3H, s), 3.38 (2H, q, J = 6.7 Hz), 3.48-3.62 (1H, m),3.90 (3H, s), 3.91 (3H, s), 4.19-4.29 (2H, m), 4.39 (1H, br s),4.72-4.83 (1H, m), 5.18 (1H, br s), 6.67-6.70 (1H, m), 7.44 (1H, s),7.57 (1H, d, J = 2.5 Hz), 7.66 (2H, s), 7.81 (2H, d, J = 8.0 Hz), 8.06(1H, d, J = 2.5 Hz), 9.21-9.36 (2H, m). MS (m/z): 603 (M + H)⁺.

TABLE 2-26 Ex. 74

Reference Example D-69

Reference Example C-14 Step 2(1R,2S,4R)-4-({[3-fluoro-4-(6-fluoro-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.79-1.86 (1H, m), 2.28 2.35 (1H, m), 2.41-2.48 (2H,m), 3.45 (3H, s), 3.56-3.62 (1H, m), 3.96 (3H, s), 4.09 (2H, q, J = 11.0Hz), 4.25-4,32 (2H, m), 4.34-4.38 (1H, m), 4.90-4.98 (1H, m), 5.21 (1H,br s), 7.55-7.58 (1H, m), 7.67 (1H, d, J = 11.0 Hz), 7.73-7.79 (2H, m),7.83 (1H, d, J = 11.0 Hz), 7.93 7.96 (1H, m), 8.36 (1H, s), 9.40-9.57(2H, m). MS (m/z): 580 (M + H)⁺. Ex. 75

Reference Example D-70

Reference Example C-20 Step 3(1R,2S,4R)-4-{[(3′-fluoro-5′-methoxy[1,1′-biphenyl]-4-yl)methyl}amino}-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.76-1.87 (1H, m), 2.20-2.47 (3H, m), 2.74-2.82 (3H,m), 3.36 (3H, s), 3.48-3.62 (1H, m), 3.81-3.97 (2H, m), 3.85 (3H, s),4.17-4.31 (2H, m), 4.34 4.43 (1H, m), 4.72- 4.85 (1H, m), 5.11-5.22 (1H,m), 6.63-6.84 (1H, m), 6.84-6.90 (1H, m), 7.09-7.13 (1H, m), 7.13- 7.18(1H, m), 7.45 (1H, s), 7.64-7.70 (2H, m), 7.78-7.84 (2H, m), 9.27-9.54(2H, m), MS (m/z): 590 (M + H)⁺.

TABLE 2-27 Ex. 76

Reference Example D-74 Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)-2-fluorophenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.77-1.86 (1H, m), 2.29-2.37 (1H, m), 2.39-2.49 (2H,m), 3.44 (3H, s), 3.62-3.70 (1H, m), 3.90 (3H, s), 3.92 (3H, s), 4.09(2H, q, J = 11.0 Hz), 4.28 (2H, t, J = 5.5 Hz), 4.36 (1H, br s),4.92-5.01 (1H, m), 5.23 (1H, br s), 7.63 (1H, d, J = 1.8 Hz), 7.70 (1H,dd, J = 8.0, 1.8 Hz), 7.76 (2H, s), 7.77-7.80 (1H, m), 8.13 (1H, d, J =2.5 Hz), 8.37 (1H, s), 9.38-9.56 (2H, m). MS (m/z): 592 (M + H)⁺. Ex. 77

Reference Example D-73 Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)-2-methoxyphenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.84 (1H, ddd, J = 14.1, 6.7, 3.1 Hz), 2.27-2.36(1H, m), 2.41-2.49 (2H, m), 3.45 (3H, s), 3.57-3.66 (1H, m), 3.91 (3H,s), 3.91 (3H, s), 3.98 (3H, s), 4.09 (2H, q, J = 11.0 Hz), 4.19 (2H, t,J = 5.5 Hz), 4.36 (1H, br s), 4.91-4.99 (1H, m), 5.26 (1H, br s), 7.35(1H, dd, J = 8.0, 1.2 Hz), 7.37 (1H, d, J = 1.2 Hz), 7.58 (1H, d, J =8.0 Hz), 7.58 (1H, s), 7.76 (1H, s), 8.10 (1H, d, J = 1.8 Hz), 8.37 (1H,s), 9.11-9.24 (2H, m). MS (m/z): 604 (M + H)⁺.

TABLE 2-28 Ex. 78

Reference Example D-72 Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)-2-methylphenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-1.90 (1H, m), 2.33-2.39 (1H, m), 2.41-2.48 (1H,m), 2.50 (3H, s), 2.52-2.60 (1H, m), 3.45 (3H, s), 3.72 (1H, br s), 3.90(3H, s), 3.91 (3H, s), 4.10 (2H, q, J = 11.0 Hz), 4.22 (2H, t, J = 6.1Hz), 4.35-4.40 (1H, m), 4.94-5.02 (1H, m), 5.10-5.27 (1H, m), 7.57 (1H,d, J = 1.8 Hz), 7.63 (2H, s), 7.65 (1H, s), 7.77 (1H, s), 8.05 (1H, d, J= 1.8 Hz), 8.38 (1H, s), 9.22- 9.32 (1H, m), 9.38-9.44 (1H, m), MS(m/z): 588 (M + H)⁺. Ex. 79

CAS:90514-72-0 Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(1H-benzimidazol-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-1.93 (1H, m), 2.28-2.38 (1H, m), 2.43-2.57 (2H,m), 3.45 (3H, s), 3.56-3.71 (1H, m), 4.10 (2H, q, J = 11.0 Hz),4.25-4.42 (3H, m), 4.91-5.03 (1H, m), 5.21 (1H, br s), 7.32-7.43 (2H,m), 7.66 (1H, d, J = 7.4 Hz), 7.73-7.93 (6H, m), 8.37 (1H, s), 8.70 (1H,s), 9.56 (1H, br s), 9.62 (1H, br s). MS: m/z 553 (M + H)⁺.

TABLE 2-29 Ex. 80

Reference Example D-48

Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(5-ethoxy-6-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.35-1.40 (3H, m), 1.78-1.90 (1H, m), 2.26-2.37 (1H,m), 2.40-2.55 (2H, m), 3.44 (3H, s), 3.51-3.66 (1H, m), 3.91 (3H, s),4.02-4.29 (6H, m), 4.31-4.39 (1H, m), 4.89-5.02 (1H, m), 5.04-5.34 (1H,m), 7.54-7.57 (1H, m), 7.65-7.70 (2H, m), 7.74-7.81 (3H, m), 8.04-8.07(1H, m), 8.36-8.38 (1H, m), 9.35-9.57 (2H, m), MS (m/z): 588 (M + H)⁺.

TABLE 2-30 Ex. 81

Reference Example D-56

Reference Example C-15 Step 2(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.84-1.97 (2H, m), 2.03-2.08 (1H, m), 2.32-2.35 (1H,m), 3.38-3.41 (2H, m), 3.52 (3H, s), 3.55-3.60 (2H, m), 3.88-3.88 (2H,m), 3.94 (6H, s), 4.06 (3H, s), 4.48-4.51 (1H, m), 5.04-5.13 (1H, m),7.25-7.29 (2H, m), 7.40 (2H, d, J = 8.0 Hz), 7.52 (2H, d, J = 8.0 Hz),7.95- 7.95 (1H, m), MS (m/z): 604 (M + H)⁺.(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.85-1.88 (1H, m), 2.29-2.33 (1H, m), 2.42-2.45 (1H,m), 2.47-2.48 (1H, m), 3.41 (3H, s), 3.52-3.55 (1H, m), 3.84 (3H, s),3.89-4.00 (8H, m), 4.22-4.24 (2H, m), 4.36-4.38 (1H, m), 4.82-4.88 (1H,m), 7.51-7.56 (2H, m), 7.67-7.77 (4H, m), 8.04 (1H, s), 9.38-9.44 (2H,m). MS (m/z): 604 (M + H)⁺.

TABLE 2-31 Ex. 82

Reference Example D-37

Reference Example C-20 Step 3(1R,2S,4R)-4-{[(3′,4′-difluoro-5′-methoxy[1,1′-biphenyl]-4-yl)methyl]amino}-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-1.84 (1H, m), 2.30-2.43 (3H, m), 2.80 (3H, s),3.37 (3H, s), 3.56- 3.59 (1H, m), 3.85-3.87 (2H, m), 3.99 (3H, s),4.24-4.26 (2H, m), 4.39-4.41 (1H, m), 4.79-4.80 (1H, m), 7.30-7.33 (2H,m), 7.41 (1H, s), 7.65 (2H, d, J = 8.3 Hz), 7.81 (2H, d, J = 8.3 Hz),9.17- 9.22 (2H, m). MS (m/z): 608 (M + H)⁺. Ex. 83

(1R,2S,4R)-4-({[4-(isoquinolin-4-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-1.92 (1H, m), 2.30-2.49 (2H, m), 2.52-2.59 (1H,m), 3.46 (3H, s), 3.66 (1H, br s), 4.10 (2H, q, J = 11.2 Hz), 4.33 (2H,br s), 4.38 (1H, br s), 4.93-5.02 (1H, m), 5.16-5.26 (1H, m), 7.67 (2H,d, J = 8.0 Hz), 7.76-7.83 (4H, m), 7.85-7.90 (2H, m), 8.30 (1H, d, J =8.6 Hz), 8.37 (1H, s), 8.49 (1H, s), 9.39-9.55 (2H, m), 9.45 (1H, s). MS(m/z): 564 (M + H)⁺.

TABLE 2-32 Ex. 84

Reference Example D-28 Reference Example C-14 Step 2(1R,2S,4R)-4-({[4-(6-chloro-5-methoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.85-1.92 (1H, m), 1.94-2.04 (1H, m), 2.06-2.13 (1H,m), 2.35 (1H, ddd, J = 14.9, 8.4, 5.4 Hz), 3.41-3.48 (1H, m), 3.54 (3H,s), 3.63 (2H, q, J = 10.2 Hz), 3.91 (1H, d, J = 13.5 Hz), 3.95 (1H, d, J= 13.5 Hz), 4.07 (3H, s), 4.48-4.52 (1H, m), 5.08 (1H, td, J = 9.7, 4.5Hz), 7.22 (1H, s), 7.40 (1H, s), 7.46-7.51 (2H, m), 7.98-8.03 (2H, m),8.40 (1H, s). MS (m/z): 579, 581 (M + H)⁺.(1R,2S,4R)-4-({[4-(6-chloro-5-methoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.80-1.90 (1H, m), 2.26-2.36 (1H, m), 2.42-2.54 (2H,m), 3.44 (3H, s), 3.55-3.67 (1H, m), 4.09 (2H, q, J = 10.7 Hz), 4.13(3H, s), 4.25-4.40 (3H, m), 4.90-5.00 (1H, m), 5.20 (1H, br s), 7.75(1H, s), 7.79 (2H, d, J = 8.0 Hz), 7.92 (1H, s), 8.28 (2H, d, J = 8.0Hz) 8.36 (1H, s), 9.47 (1H, br s), 9.54 (1H, br s). MS (m/z): 579, 581(M + H)⁺.

TABLE 2-33 Ex. 85

Reference Example C-14 Step 3 Reference Example D-41(1R,2S,4R)-4-[({4-[6-fluoro-1-(methanesulfonyl)-1H-indazol-4-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.79-1.88 (1H, m), 2.29-2.36 (1H, m), 2.38-2.49 (2H,m), 3.45 (3H, s), 3.57 (3H, s), 3.60-3.68 (1H, m), 4.10 (2H, q, J = 11.2Hz), 4.31 (2H, br s), 4.38 (1H, br s), 4.92- 5.01 (1H, m), 5.21 (1H, brs), 7.54 (1H, dd, J = 10.1, 2.1 Hz), 7.74 (1H, dd, J = 8.6, 2.5 Hz),7.76 (1H, s), 7.78 (2H, d, J = 8.0 Hz), 7.88 (2H, d, J = 8.0 Hz), 8.37(1H, s), 8.65 (1H, d, J = 1.2 Hz), 9.30-9.45 (2H, m). MS (m/z): 649 (M +H)⁺. Ex. 86

Reference Example D-44 Reference Example C-14 Step 3(1R,2S,4R)-4-[({3′-[(methanesulfonyl)methyl][1,1′biphenyl]-4-yl}methyl)amino]-2-{methyl[6-(2,2,2 trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.79-1.83 (1H, m), 2.27-2.47 (3H, m), 2.95 (3H, s),3.44 (3H, s), 3.58- 3.61 (1H, m), 4.09 (2H, q, J = 11.0 Hz), 4.23-4.26(2H, m), 4.34-4.37 (1H, m), 4.58 (2H, s), 4.94- 4.95 (1H, m), 5.18-5.21(1H, m), 7.43-7.77 (9H, m), 8.36 (1H, s), 9.32-9.34 (2H, m). MS (m/z):605 (M + H)⁺.

TABLE 2-34 Ex. 87

Reference Example D-33

Reference Example C-20 Step 3(1R,2S,4R)-4-{[(2′,4′-difluoro-3′-methoxy[1,1′-biphenyl]-4-yl)methyl]amino}-2-{methyl[2-(methylamino)-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.82-1.86 (1H, m), 2.27-2.33 (1H, m), 2.39-2.48 (2H,m), 2.81 (3H, s), 3.38 (3H, s), 3.55-3.58 (1H, m), 3.90-3.96 (5H, m),4.24-4.26 (2H, m), 4.38-4.40 (1H, m), 4.79- 4.82 (1H, m), 5.17-5.20 (1H,m), 7.23-7.30 (2H, m), 7.49 (1H, s), 7.60-7.62 (2H, m), 7.70-7.72 (2H,m), 9.46-9.52 (2H, m). MS (m/z): 608 (M + H)⁺. Ex. 88

Reference Example C-14 Step 3 Reference Example D-47(1R,2S,4R)-4-[({4-[5-methoxy-6-(trifluoromethyl)pyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-1.90 (1H, m), 2.26 2.36 (1H, m), 2.40 2.55 (2H,m), 3.45 (3H, s), 3.53-3.65 (1H, m), 4.05 (3H, s), 4.05-4.15 (2H, m),4.17-4.32 (2H, m), 4.32-4.38 (1H, m), 4.90- 5.00 (1H, m), 5.05-5.39 (1H,m), 7.70 7.78 (3H, m), 7.84 7.90 (2H, m), 8.36 8.40 (2H, m), 8.81- 8.85(1H, m), 9.41-9.62 (2H, m). MS (m/z): 612 (M + H)⁺.

TABLE 2-35 Ex. 89

Reference Example D-26

Reference Example C-24 Step 22-[(4-{[(1S,2R,4R)-4({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-5-fluoro-N,N-di(propan-2-yl)benzamidehydrochloride

¹H-NMR (DMSO-D₆) δ: 0.99-1.14 (6H, m), 1.24-1.48 (5H, m), 1.68-1.80 (1H,m), 2.06-2.23 (1H, m), 2.30-2.55 (2H, m), 3.22 (3H, s), 3.27-3.61 (3H,m), 3.68-3.80 (1H, m), 4.01 (3H, s), 4.07 (3H, s), 4.08-4.32 (3H, m),4.48-4.66 (1H, m), 5.12-5.23 (1H, m), 6.79-6.91 (1H, m), 7.16-7.26 (2H,m), 7.64-7.73 (3H, m), 7.94-7.99 (1H, m), 8.15-8.22 (2H, m), 8.46 (1H,s), 9.20-9.47 (2H, m). MS (m/z): 674 (M + H)⁺. Ex. 90

Reference Example D-28

Reference Example C-15 Step 2(1R,2S,4R)-4-({[4-(6-chloro-5methoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.86-1.89 (1H, m), 2.29-2.35 (2H, m), 2.43-2.50 (2H,m), 2.50-2.53 (1H, m), 3.42 (3H, s), 3.85 (3H, s), 3.96 (2H, q, J = 11.0Hz), 4.13 (3H, s), 4.28 4.31 (2H, m), 4.37- 4.39 (1H, m), 4.82-4.88 (1H,m), 7.59 (1H, s), 7.78 (2H, d, J = 8.6 Hz), 7.87 (1H, s), 8.25 (2H, d, J= 8.6 Hz), 9.44-9.48 (2H, m). MS (m/z): 609, 611 (M + H)⁺.

TABLE 2-36 Ex. 91

Reference Example D-68

Reference Example C-15 Step 2(1R,2S,4R)-4-({[4-(6-fluoro-5-methoxypyridin-3yl)phenyl]methyl}amino)-2{[2-methoxy6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.86-1.89 (1H, m), 2.30-2.33 (1H, m), 2.43-2.46 (2H,m), 3.42 (3H, s), 3.59-3.68 (1H, m), 3.85 (3H, s), 3.93-3.98 (5H, m),4.24-4.26 (2H, m), 4.37-4.38 (1H, m), 4.82- 4.88 (1H, m), 7.59 (1H, s),7.72 (2H, d, J = 8.6 Hz), 7.84 (2H, d, J = 8.6 Hz), 7.88-7.90 (1H, m),8.05-8.06 (1H, m), 9.43-9.47 (2H, m). MS (m/z): 592 (M + H)⁺. Ex. 92

Reference Example C-14 Step 3 Reference Example D-79 Step 1(1R,2S,4R)-4-[({4-[5-methoxy-6-(methylamino)pyridin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.79-1.88 (1H, m), 2.25-2.36 (1H, m), 2.37-2.53 (2H,m), 2.85-2.91 (3H, m), 3.44 (3H, s), 3.50-3.64 (1H, m), 3.91 (3H, s),4.00 4.30 (4H, m), 4.32-4.39 (1H, m), 4.88 4.99 (1H, m), 5.17-5.23 (1H,m), 6.18-6.74 (1H, m), 7.31-7.35 (1H, m), 7.59-7.66 (2H, m), 7.70-7.77(3H, m), 7.96-8.00 (1H, m), 8.36 (1H, s), 9.31-9.53 (2H, m). MS (m/z):573 (M + H)⁺.

TABLE 2-37 Ex. 93

Reference Example D-26 Reference Example C-21 Step 2(1R,2S,4R)-2-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)cyclopentan-1-ol

¹H-NMR (DMSO-D₆) δ: 1.49-1.52 (1H, m), 1.97-2.05 (1H, m), 2.12-2.18 (1H,m), 2.23-2.30 (1H, m), 3.08-3.10 (1H, m), 3.41 (3H, s), 3.813.84 (2H,m), 3.99 (3H, s), 4.00 4.05 (2H, m), 4.06 (3H, s), 4.29-4.31 (1H, m),4.67-4.73 (2H, m), 7.49 (2H, d, J = 8.0 Hz), 7.55 (1H, s), 7.68 (1H, s),8.02 (2H, d, J = 8.0 Hz). MS (m/z): 609, 611 (M + H)⁺.(1R,2S,4R)-2-{[2-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.86-1.90 (1H, m), 2.37-2.44 (2H, m), 2.51-2.54 (1H,m), 3.44 (3H, s), 3.61-3.63 (1H, m), 4.01-4.10 (8H, m), 4.26-4.28 (2H,m), 4.36-4.38 (1H, m), 4.79-4.85 (1H, m), 7.63 (1H, s), 7.73 (3H, d, J =8.6 Hz), 8.17 (2H, d, J = 8.6 Hz), 9.41-9.48 (2H, m). MS (m/z): 609, 611(M + H)⁺.

TABLE 2-38 Ex. 94

Reference Example D-52

Reference Example C-15 Step 2(1R,2S,4R)-4-[({4-[6-(methanesulfonyl)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.75-1.88 (1H, m), 2.25-2.48 (3H, m), 3.34 (3H, s),3.41 (3H, s), 3.55- 3.64 (1H, m), 3.84 (3H, s), 4.00 (2H, q, J = 11.0Hz), 4.09 (3H, s), 4.29 (2H, br s), 4.37 (1H, br s), 4.84 (1H, br s),5.20 (1H, br s), 7.64 (1H, s), 7.73-7.80 (2H, m), 7.99 (2H, d, J = 8.0Hz), 8.05 (1H, d, J = 1.8 Hz), 8.61 (1H, d, J = 1.8 Hz), 9.27-9.49 (2H,m). MS (m/z): 652 (M + H)⁺.

TABLE 2-39 Ex. 95

Reference Example C-4 Step 2 Reference Example D-26(1R,3S)-N³-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine

¹H-NMR (CDCl₃) δ: 1.51-1.60 (1H, m), 1.65-1.71 (1H, m), 1.90-2.00 (3H,m), 2.26-2.33 (1H, m), 3.24-3.31 (1H, m), 3.32 (3H, s), 3.63 (2H, q, J =10.0 Hz), 3.84-3.90 (2H, m), 4.03 (3H, s), 4.24 (3H, s), 5.30-5.39 (1H,m), 7.14 (1H, s), 7.35 (1H, s), 7.46 (2H, d, J = 8.6 Hz), 7.97 (2H, d, J= 8.6 Hz), 8.42 (1H, s).(1R,3S)-N³-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-(trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

¹H-NMR (DMSO-D₆) δ: 1.88-2.15 (5H, m), 2.33-2.41 (1H, m), 3.29 (3H, s),3.61-3.70 (1H, m), 4.01 (3H, s), 4.05-4.15 (5H, m), 4.24-4.29 (2H, m),5.25-5.32 (1H, m), 7.67 (1H, s), 7.71-7.74 (3H, m), 8.19-8.22 (2H, m),8.38 (1H, s), 9.25-9.48 (2H, m). MS (m/z): 559 (M + H)⁺.

TABLE 2-40 Ex. 96

(1R,2S,4R)-4-({[4-(5,6-dimethoxypyrazin-2-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.70-1.82 (1H, m), 2.22-2.43 (2H, m), 3.43 (3H, s),3.57 (1H, br s), 3.95 (3H, s), 4.00-4.15 (5H, m), 4.24 (2H, br s),4.31-4.40 (2H, m), 4.87-4.99 (1H, m), 5.20 (1H, br s), 7.65 (2H, d, J =8.5 Hz), 7.75 (1H, s), 8.10 (2H, d, J = 8.5 Hz), 8.36 (1H, s), 8.38 (1H,s), 9.24 (2H, br s). MS (m/z): 575 (M + H)⁺. Ex. 97

(1R,2S,4R)-4-({[4-(4,5-dimethoxypyrimidin-2-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.74-1.81 (1H, m), 2.24-2.44 (3H, m), 3.44 (3H, s),3.55-3.65 (1H, m), 3.92 (3H, s), 4.08 (3H, s), 4.09 (2H, q, J = 10.4Hz), 4.27 (2H, br s), 4.37 (1H, br s), 4.89-4.98 (1H, m), 5.17-5.22 (1H,m), 7.67 (2H, d, J = 8.0 Hz), 7.75 (1H, s), 8.36 (2H, s), 8.36 (2H, d, J= 8.0 Hz), 9.15-9.27 (2H, m). MS (m/z): 575 (M + H)⁺.

TABLE 2-41 Ex. 98

(1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.73-1.84 (1H, m), 2.23-2.46 (3H, m), 3.08 (6H, s),3.44 (3H, s), 3.57- 3.68 (1H, m), 4.03 (3H, s), 4.09 (2H, q, J = 11.0Hz), 4.28 (2H, br s), 4.37 (1H, br s), 4.90-4.99 (1H, m), 5.21 (1H, brs), 7.54 (1H, s), 7.68-7.73 (2H, m), 7.75 (1H, s), 8.17 (2H, d, J = 8.6Hz), 8.36 (1H, s), 9.15-9.33 (2H, m). MS (m/z): 588 (M + H)⁺. Ex. 99

6-[4-({[(1R,3R,4S)-3-hydroxy-4-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentyl]amino}methyl)phenyl]-4-methoxypyridazine-3-carbonitrilehydrochloride

¹H-NMR (DMSO-D₆) δ: 1.78-1.86 (1H, m), 2.27-2.34 (1H, m), 2.41-2.49 (2H,m), 3.44 (3H, s), 3.59-3.67 (1H, m), 4.09 (2H, q, J = 10.4 Hz), 4.19(3H, s), 4.29-4.38 (3H, m), 4.91-4.99 (1H, m), 5.18-5.24 (1H, m), 7.75(1H, s), 7.81 (2H, d, J = 8.0 Hz), 8.13 (1H, s), 8.36 (1H, s), 8.38 (2H,d, J = 8.0 Hz), 9.38 (2H, br s). MS (m/z): 570 (M + H)⁺.

TABLE 2-42 Ex. 100

(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-[(5-{4-fluoro-2-[4-(propan-2-yl)pyrimidin-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.07-1.16 (6H, m), 1.75-1.77 (1H, m), 2.14-2.16 (1H,m), 2.31-2.34 (2H, m), 3.03-3.06 (4H, m), 4.01 (3H, s), 4.07 (3H, s),4.21-4.23 (3H, m), 4.48-4.51 (1H, m), 5.10-5.13 (1H, m), 7.00-7.03 (1H,m), 7.34-7.36 (1H, m), 7.42-7.44 (1H, m), 7.66 (1H, s), 7.71 (2H, d, J =8.6 Hz), 7.98-8.00 (1H, m), 8.18 (2H, d, J = 8.6 Hz), 8.41 (1H, s),8.67-8.72 (1H, m), 9.17 (1H, s), 9.37-9.44 (2H, m). MS (m/z): 667 (M +H)⁺. Ex. 101

(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-[(5-{[5-fluoro-2′-(propan-2-yl)[1,1′-biphenyl]-2-yl]oxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.09-1.12 (6H, m), 1.70-1.74 (1H, m), 2.10-2.13 (1H,m), 2.26-2.33 (2H, m), 2.79-2.82 (1H, m), 3.06 (3H, d, J = 6.1 Hz), 4.00(3H, s), 4.07 (3H, s), 4.21-4.24 (3H, m), 4.40-4.47 (1H, m), 5.10-5.12(1H, m), 6.94-6.97 (1H, m), 7.13-7.28 (4H, m), 7.34-7.40 (2H, m), 7.66(1H, s), 7.70 (2H, d, J = 8.0 Hz), 7.86 (1H, d, J = 10.4 Hz), 8.19 (2H,d, J = 8.0 Hz), 8.37 (1H, d, J = 4.3 Hz), 9.30-9.33 (2H, m). MS (m/z):665 (M + H)⁺.

TABLE 2-43 Ex. 102

(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-[(5-{4-fluoro-2-[1-(propan-2-yl)-1H-pyrazol-5-yl]phenoxy}pyrimidin-4-yl)(methyl)amino]cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.31-1.34 (6H, m), 1.70-1.75 (1H, m), 2.04-2.17 (2H,m), 2.26-2.34 (2H, m), 3.08 (3H, s), 4.01 (3H, s), 4.07 (3H, s),4.13-4.16 (1H, m), 4.22 (2H, s), 4.40-4.45 (2H, m), 5.13-5.16 (1H, m),6.29 (1H, s), 6.98-6.99 (1H, m), 7.34-7.36 (2H, m), 7.53 (1H, s), 7.66(1H, s), 7.70 (2H, d, J = 8.6 Hz), 8.06 (1H, s), 8.18 (2H, d, J = 8.6Hz), 8.46 (1H, s), 9.33-9.39 (2H, m). MS (m/z): 655 (M + H)⁺. Ex. 103

(1S,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.93-2.05 (2H, m), 2.24-2.32 (1H, m), 2.38-2.44 (1H,m), 3.30 (3H, s), 3.71-3.81 (1H, m), 4.00 (3H, s), 4.01-4.07 (2H, m),4.08 (3H, s), 4.18-4.25 (2H, m), 4.49-4.56 (1H, m), 4.89-4.97 (1H, m),5.01-5.13 (1H, m), 7.59 (1H, s), 7.71 (2H, d, J = 8.0 Hz), 7.72 (1H, s),8.15 (2H, d, J = 8.0 Hz), 8.35 (1H, s), 9.32 (2H, br s). MS (m/z): 575(M + H)⁺.

TABLE 2-44 Ex. 104

6-[(4-{[(1S,2R,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-hydroxycyclopentyl](methyl)amino}pyrimidin-5-yl)oxy]-2,3-difluoro-N,N-di(propan-2-yl)benzamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.04-1.45 (12H, m), 1.72-1.79 (1H, m), 2.04-2.20(1H, m), 2.33-2.46 (2H, m), 3.19 (3H, s), 3.47-3.49 (2H, m), 3.59-3.64(1H, m), 3.83-3.90 (1H, m), 4.00 (3H, s), 4.07 (3H, s), 4.24 (2H, s),4.50-4.60 (1H, m), 5.16-5.18 (1H, m), 6.61-6.66 (1H, m), 7.40-7.45 (1H,m), 7.66 (1H, s), 7.69-7.72 (2H, m), 8.00-8.02 (1H, m), 8.16-8.18 (2H,m), 8.47 (1H, s), 9.40-9.42 (2H, m). MS (m/z): 692 (M + H)⁺. Ex. 105

(1R,2S,4R)-4-[({4-[6-(azetidin-1-yl)-5-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.85-1.91 (1H, m), 2.29-2.37 (3H, m), 2.43-2.47 (2H,m), 3.44 (3H, s), 3.58-3.66 (1H, m), 3.93 (3H, s), 4.02 (2H, q, J = 10.9Hz), 4.19 (4H, t, J = 7.3 Hz), 4.23-4.25 (2H, m), 4.36-4.40 (1H, m),4.91-4.98 (1H, m), 5.02 (1H, br s), 7.35 (1H, s), 7.65-7.68 (3H, m),8.09 (2H, d, J = 8.5 Hz), 8.33 (1H, s), 9.24 (2H, br s). MS (m/z): 600(M + H)⁺.

TABLE 2-45 Ex. 106

5-[4-({[(1R,3R,4S)-3-hydroxy-4-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]amino}methyl)phenyl]-3-methoxypyridine-2-carbonitrilehydrochloride

¹H-NMR (DMSO-D₆, 80° C.) δ: 1.83-1.90 (1H, m), 2.29-2.36 (1H, m),2.39-2.47 (2H, m), 3.42 (3H, s), 3.58-3.65 (1H, m), 3.85 (3H, s), 3.94(2H, q, J = 11.0 Hz), 4.09 (3H, s), 4.24-4.31 (2H, m), 4.37-4.41 (1H,m), 4.81-4.88 (1H, m), 4.98-5.10 (1H, m), 7.57 (1H, s), 7.75 (2H, d, J =8.6 Hz), 7.95 (2H, d, J = 8.6 Hz), 7.98 (1H, d, J = 1.8 Hz), 8.66 (1H,d, J = 1.8 Hz), 9.30 (2H, br s). MS (m/z): 599 (M + H)⁺. Ex. 107

(1R,2S,4R)-4-{[(4-{5-methoxy-6-[(pyridin-3-yl)oxy]pyridazin-3-yl}phenyl)methyl]amino}-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.82-1.88 (1H, m), 2.28-2.32 (1H, m), 2.40-2.45 (2H,m), 3.44 (3H, s), 3.60-3.63 (1H, m), 4.06-4.11 (5H, m), 4.27-4.29 (2H,m), 4.35-4.37 (1H, m), 4.93-4.95 (1H, m), 5.19-5.21 (1H, m), 7.53-7.57(1H, m), 7.74-7.78 (4H, m), 7.90 (1H, s), 8.21 (2H, d, J = 8.6 Hz), 8.36(1H, s), 8.50-8.51 (1H, m), 8.57-8.57 (1H, m), 9.35-9.42 (2H, m). MS(m/z): 638 (M + H)⁺.

TABLE 2-46 Ex. 108

(1R,2S,4R)-4-{[(4-{5-methoxy-6-[(oxan-3-yl)oxy]pyridazin-3-yl}phenyl)methyl]amino}-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride (diastereomer mixture)

¹H-NMR (DMSO-D₆) δ: 1.57-1.59 (1H, m), 1.81-1.91 (3H, m), 2.10-2.14 (1H,m), 2.29-2.35 (1H, m), 2.44-2.46 (1H, m), 2.52-2.53 (1H, m), 3.45 (3H,s), 3.54-3.69 (4H, m), 3.91-3.94 (1H, m), 4.00-4.07 (5H, m), 4.26-4.28(2H, m), 4.36-4.38 (1H, m), 4.92-4.98 (1H, m), 5.23-5.28 (1H, m), 7.63(1H, s), 7.72-7.73 (3H, m), 8.16 (2H, d, J = 8.0 Hz), 8.35 (1H, s),9.39-9.46 (2H, m). MS (m/z): 645 (M + H)⁺. Ex. 109

(1R,2S,4R)-4-[({4-[5-methoxy-6-(2-methoxyethoxy)pyridazin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.74-1.84 (1H, m), 2.25-2.46 (3H, m), 3.32 (3H, s),3.41 (3H, s), 3.57- 3.64 (1H, m), 3.72-3.75 (2H, m), 3.84 (3H, s), 4.00(2H, q, J = 10.4 Hz), 4.01 (3H, s), 4.28 (2H, br s), 4.38 (1H, br s),4.57-4.60 (2H, m), 4.80-4.87 (1H, m), 5.20 (1H, br s), 7.64 (1H, s),7.67 (1H, s), 7.70 (2H, d, J = 8.0 Hz), 8.20 (2H, d, J = 8.0 Hz),9.15-9.32 (2H, m). MS (m/z): 649 (M + H)⁺.

TABLE 2-47 Ex. 110

(1R,2S,4R)-4-[({4-[6-(difluoromethoxy)-4-methoxypyridazin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.81-2.08 (1H, m), 2.28-2.35 (1H, m), 2.42-2.45 (1H,m), 2.53-2.55 (1H, m), 3.41 (3H, s), 3.60-3.62 (1H, m), 3.85 (3H, s),3.99-4.02 (5H, m), 4.26-4.28 (2H, m), 4.37-4.38 (1H, m), 4.84-4.86 (1H,m), 7.28 (1H, s), 7.78-8.01 (6H, m), 9.48-9.54 (2H, m). MS (m/z): 641(M + H)⁺. Ex. 111

2-(difluoromethyl)-6-[4-({[(1R,3R,4S)-3-hydroxy-4-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]amino}methyl)phenyl]-5-methoxypyridazin-3(2H)-one hydrochloride

¹H-NMR (DMSO-D₆) δ:1.77-2.45 (4H, m), 3.41 (3H, s), 3.59-3.61 (1H, m),3.84 (3H, s), 3.89 (3H, s), 3.98-4.01 (2H, m), 4.25-4.28 (2H, m),4.36-4.39 (1H, m), 4.82-4.85 (1H, m), 5.18-5.20 (1H, m), 6.55 (1H, s),7.63-7.73 (5H, m), 7.97 (1H, t, J = 58.3 Hz), 9.31-9.38 (2H, m). MS(m/z): 641 (M + H)⁺.

TABLE 2-48 Ex. 112

(1R,2S,4R)-4-[({4-[6-(hydroxymethyl)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆, 80° C.) δ: 1.84-1.91 (1H, m), 2.28-2.37 (1H, m),2.39-2.50 (2H, m), 3.42 (3H, s), 3.56-3.68 (1H, m), 3.85 (3H, s), 3.94(2H, q, J = 10.6 Hz), 3.94 (3H, s), 4.22-4.30 (2H, m), 4.36-4.42 (1H,m), 4.60 (2H, s), 4.81-4.88 (1H, m), 5.00-5.10 (1H, m), 7.57 (1H, s),7.63 (1H, d, J = 1.8 Hz), 7.70 (2H, d, J = 7.4 Hz), 7.84 (2H, d, J = 8.6Hz), 8.44 (1H, d, J = 1.8 Hz), 9.30 (2H, s). MS (m/z): 604 (M + H)⁺. Ex.113

N-{6-[4-({[(1R,3R,4S)-3-hydroxy-4-{[2-methoxy-6(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentyl]amino}methyl)phenyl]-4-methoxypyridazin-3-yl}-N-methylprop-2-enamide hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-2.47 (4H, m), 3.42 (3H, s), 3.59-3.61 (1H, m),3.85 (3H, s), 3.87 (3H, s), 3.94-3.97 (5H, m), 4.25-4.28 (2H, m),4.36-4.38 (1H, m), 4.81-4.87 (1H, m), 5.09-5.12 (1H, m), 5.62 (1H, dd, J= 10.4, 1.8 Hz), 6.01 (1H, dd, J = 17.2, 1.8 Hz), 6.27 (1H, dd, J =17.2, 10.4 Hz), 7.41 (1H, s), 7.59 (1H, s), 7.73 (2H, d, J = 8.3 Hz),8.05 (2H, d, J = 8.3 Hz), 9.44-9.47 (2H, m). MS (m/z): 658 (M + H)⁺.

TABLE 2-49 Ex. 114

(1R,2S,4R)-4-({[4-(5-methoxy-6-phenoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethy)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-2.46 (4H, m), 3.41 (3H, s), 3.58-3.61 (1H, m),3.84 (3H, s), 3.98- 4.01 (2H, m), 4.10 (3H, s), 4.27 (2H, s), 4.35-4.37(1H, m), 4.83-4.84 (1H, m), 7.20-7.29 (3H, m), 7.44-7.49 (2H, m), 7.64(1H, s), 7.76-7.78 (2H, m), 8.20 (2H, d, J = 8.6 Hz), 9.57-9.63 (2H, m).MS (m/z): 667 (M + H)⁺. Ex. 115

(1R,2S,4R)-4-({[1-(5,6-dimethoxypyridazin-3-yl)piperidin-4-yl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.27-1.30 (2H, m), 1.74-2.47 (7H, m), 2.87-2.89 (4H,m), 3.40 (3H, s), 3.53-3.58 (1H, m), 3.84 (3H, s), 3.88 (3H, s), 3.89(3H, s), 3.98-4.01 (2H, m), 4.22-4.25 (2H, m), 4.35-4.36 (1H, m),4.81-4.82 (1H, m), 5.16-5.17 (1H, m), 6.86 (1H, s), 7.63 (1H, s),8.93-9.02 (2H, m). MS (m/z): 612 (M + H)⁺.

TABLE 2-50 Ex. 116

(1R,2S,4R)-4-({[4-(4,5-dimethoxypyridin-2-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.79-2.08 (1H, m), 2.27-2.33 (1H, m), 2.40-2.46 (2H,m), 3.41 (3H, s), 3.58-3.60 (1H, m), 3.84 (3H, s), 3.91 (3H, s),3.98-4.01 (5H, m), 4.24-4.26 (2H, m), 4.35-4.38 (1H, m), 4.82-4.85 (1H,m), 5.17-5.20 (1H, m), 7.62 (1H, s), 7.63 (1H, s), 7.69 (2H, d, J = 8.0Hz), 8.15 (2H, d, J = 8.0 Hz), 8.27 (1H, s), 9.38-9.44 (2H, m). MS(m/z): 604 (M + H)⁺. Ex. 117

(1R,2S,4R)-4-[({4-[6-(dimethylamino)-5-methoxypyridin-3-yl]phenyl}methyl)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.83-2.47 (4H, m), 2.99 (6H, s), 3.42 (3H, s),3.58-3.61 (1H, m), 3.85 (3H, s), 3.90 (3H, s), 3.94-3.97 (2H, m),4.21-4.23 (2H, m), 4.36-4.38 (1H, m), 4.81-4.88 (1H, m), 7.46 (1H, d, J= 1.8 Hz), 7.59 (1H, s), 7.65 (2H, d, J = 8.3 Hz), 7.75 (2H, d, J = 8.3Hz), 8.10 (1H, d, J = 1.8 Hz), 9.36-9.42 (2H, m). MS (m/z): 617 (M +H)⁺.

TABLE 2-51 Ex. 118

(1R,2S,4R)-4-({[4-(6-{[1-(methanesulfonyl)piperidin-4-yl]oxy}-5-methoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl(methyl)amino}cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆) δ: 1.80-2.42 (8H, m), 2.93 (3H, s), 3.12-3.16 (2H, m),3.36-3.47 (6H, m), 3.84 (3H, s), 3.97-4.01 (5H, m), 4.27-4.29 (2H, m),4.37-4.38 (1H, m), 4.83-4.86 (1H, m), 5.34- 5.40 (1H, m), 7.64 (1H, s),7.70 (1H, s), 7.74 (2H, d, J = 8.3 Hz), 8.19 (2H, d, J = 8.3 Hz), 9.39-9.46 (2H, m) MS (m/z): 752 (M + H)⁺. Ex. 119

(1R,2S,4R)-4-[({4-[6-(difluoromethoxy)-5-methoxypyridazin-3-yl]phenyl}methy)amino]-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.73-1.83 (1H, m), 2.28-2.44 (2H, m), 3.42 (3H, s),3.62 (1H, s), 3.84 (3H, s), 4.00 (2H, dd, J = 22.0, 11.0 Hz), 4.09 (3H,s), 4.30 (2H, s), 4.39 (1H, s), 4.84 (1H, br s), 5.20 (1H, d, J = 4.3Hz), 7.63 (1H, s), 7.69-8.10 (4H, m), 8.24 (2H, d, J = 8.5 Hz), 9.27(2H, br s). MS (m/z): 641 (M + H)⁺.

TABLE 2-52 Ex. 120

(1R,2S,4R)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}-4-[({3-[(pyridin-3-yl)amino]phenyl}methyl)amino]cyclopentan-1-olhydrochloride

¹H-NMR (DMSO-D₆) δ: 1.74-1.75 (1H, m), 2.04-2.08 (2H, m), 2.27-2.36 (1H,m), 3.40 (3H, s), 3.59 (1H, s), 3.84 (3H, s), 3.99 (2H, q, J = 11.0 Hz),4.15 (2H, s), 4.37 (1H, s), 4.82 (1H, s), 5.18 (1H, s), 7.02-7.13 (2H,m), 7.29-7.40 (3H, m), 7.53-7.65 (2H, m), 8.10 (1H, d, J = 4.9 Hz), 8.46(1H, d, J = 2.4 Hz), 9.15 (2H, br s). MS (m/z): 559 (M + H)⁺. Ex. 121

(1R,2S,4R)-4-{[(4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}phenyl)methyl[amino}-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.86-2.09 (3H, m), 2.34 (1H, ddd, J = 14.9, 8.2, 5.2Hz), 3.42-3.47 (1H, m), 3.54 (3H, s), 3.63 (2H, q, J = 10.3 Hz),3.87-3.95 (2H, m), 4.47-4.50 (1H, m), 5.11 (1H, td, J = 9.6, 4.7 Hz),7.13 (1H, s), 7.41 (1H, s), 7.45 (2H, d, J = 7.9 Hz), 7.98 (2H, d, J =7.9 Hz), 8.40 (1H, s). MS (m/z): 581 (M + H)⁺.

TABLE 2-53 Ex. 122

(1R,2S,4R)-4-({[4-(6-{bis[(²H₃)methyl]amino}-5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.86-2.08 (3H, m), 2.31-2.38 (1H, m), 3.42-3.48 (1H,m), 3.54 (3H, s), 3.62 (2H, q, J = 10.3 Hz), 3.83-3.91 (2H, m), 3.92(3H, s), 4.47-4.50 (1H, m), 5.13 (1H, td, J = 9.6, 4.5 Hz), 7.20 (1H, d,J = 1.9 Hz), 7.37-7.41 (3H, m), 7.53 (2H, d, J = 8.5 Hz), 8.09 (1H, d, J= 1.9 Hz), 8.39 (1H, s). MS (m/z): 593 (M + H)⁺. Ex. 123

(1R,2S,4R)-4-{[(4-{5,6-bis[(²H₃)methyloxy]pyridazin-3-yl}phenyl)methyl}amino}-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl}amino}cyclopentan-1-ol

¹H-NMR (CDCl₃) δ: 1.85-2.06 (3H, m), 2.30-2.38 (1H, m), 3.41-3.46 (1H,m), 3.52 (3H, s), 3.57 (2H, q, J = 10.3 Hz), 3.86-3.94 (2H, m), 3.96(3H, s), 4.47-4.50 (1H, m), 5.12 (1H, td, J = 9.4, 4.5 Hz), 7.13 (1H,s), 7.32 (1H, s), 7.44 (2H, d, J = 7.9 Hz), 7.98 (2H, d, J = 7.9 Hz). MS(m/z): 611(M + H)⁺.

TABLE 2-54 Ex. 124

(1R,2S,4R)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}-4-({[3-(pyridin-3-yl)phenyl]methyl}amino)cyclopentan-1-ol hydrochloride

¹H-NMR (DMSO-D₆)δ: 1.80-1.88 (1H, m), 2.27-2.35 (1H, m), 2.40-2.53 (2H,m), 3.44 (3H, s), 3.58-3.68 (1H, m), 4.06 (1H, d, J = 11.0 Hz), 4.11(1H, d, J = 11.0 Hz), 4.23-4.39 (3H, m), 4.90- 4.99 (1H, m), 5.19 (1H,s), 7.53-7.64 (3H, m), 7.75 (1H, s), 7.81 (1H, d, J = 8.0 Hz), 8.04 (1H,s), 8.16 (1H, dt, J = 8.0, 2.0 Hz), 8.36 (1H, s), 8.62 (1H, dd, J = 5.0,2.0 Hz), 8.98 (1H, d, J = 2.0 Hz), 9.33-9.56 (2H, m). MS(m/z): 514 (M +H)⁺. Ex. 125

(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)cyclohex-3-en-1-yl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride (diastereomer mixture)

¹H-NMR (DMSO-D₆)δ: 1.37-1.49 (1H, m), 1.77-1.85 (1H, m), 2.03-2.16 (4H,m), 2.22-2.31 (1H, m), 2.37-2.48 (3H, m), 2.71-2.81 (1H, m), 2.91-3.02(2H, m), 3.41(3H, s), 3.55-3.65 (1H, m), 3.85 (3H, s), 3.95-4.05 (8H,m), 4.33-4.38 (1H, m), 4.77-4.87 (1H, m), 6.76 (1H, s), 7.48 (1H, s),7.64 (1H, s), 8.93-9.29 (2H, m). MS(m/z): 609 (M + H)⁺.

Example 126(1S,3S,4S)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxy-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride Step 1(1S,3S,4S)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxy-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diamine

The title compound was obtained in the same manner as in Step 1 ofReference Example C-2, using the compound obtained in Step 2 ofReference Example E-3 and4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine producedaccording to the method described in a literature (cancer cell, 2015,27, 589-602.).

¹H-NMR (CDCl₃) δ: 1.52-1.84 (4H, m), 1.97-2.13 (2H, m), 2.17-2.26 (1H,m), 3.15-3.21 (1H, m), 3.31-3.42 (2H, m), 3.46 (3H, s), 3.48-3.52 (1H,m), 3.92 (2H, s), 4.03 (3H, s), 4.25 (3H, s), 4.56-4.64 (1H, m), 6.52(1H, s), 7.16 (1H, s), 7.47-7.51 (2H, m), 8.03-8.08 (2H, m), 8.08-8.25(1H, m), 8.43 (1H, s).

MS (m/z): 589 (M+H)⁺.

Step 2(1S,3S,4S)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxy-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclohexane-1,3-diaminehydrochloride

To a solution of the compound (0.143 g) obtained in the above Step 1 inethanol (2.00 mL) was added 5N hydrochloric acid (0.0486 mL) at roomtemperature, and the solvent was evaporated under reduced pressure, andthe residue was dried. The obtained residue was suspended in diethylether, and the resulting solid was collected by filtration, and dried togive the title compound (0.142 g).

¹H-NMR (DMSO-D6) δ: 1.19-1.34 (1H, m), 1.52-1.76 (2H, m), 2.20-2.34 (2H,m), 2.40-2.48 (1H, m), 3.26 (3H, s), 3.27-3.41 (2H, m), 4.01 (3H, s),4.05-4.37 (5H, m), 4.07 (3H, s), 7.67-7.76 (4H, m), 8.14-8.20 (2H, m),8.23-8.31 (1H, m), 8.36 (1H, s), 9.27-9.57 (2H, m).

MS (m/z): 589 (M+H)⁺.

Example 127(1R,3S,4R)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxy-N³-methyl-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride Step 1(1R,3S,4R)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxy-N³-methyl-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diamine

A mixture of the compound (0.638 g) obtained in Reference Example E-4,hydrogen chloride (4 mol/L, 1,4-dioxane solution, 13.5 mL) anddichloromethane (15.0 mL) was stirred at room temperature for 35 min,and concentrated. The residue was dried to give a solid (0.601 g). Amixture of the obtained solid,4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (0.358 g)produced according to the method described in a literature (cancer cell,2015, 27, 589-602), DIPEA (1.41 mL) and 2-propanol (20 mL) was stirredat 80° C. for 5 hr, and concentrated under reduced pressure, and theobtained residue was subjected to silica gel column chromatography(ethyl acetate/methanol to give the title compound (0.454 g) as a solid.

¹H-NMR (CDCl₃) δ: 1.67-1.74 (1H, m), 2.01-2.10 (1H, m), 2.20-2.34 (2H,m), 3.22 (3H, s), 3.22-3.28 (1H, m), 3.48 (3H, s), 3.63 (2H, q, J=10.2Hz), 3.88-3.94 (2H, m), 4.00-4.04 (1H, m), 4.03 (3H, s), 4.24 (3H, s),5.01-5.08 (1H, m), 7.14 (1H, s), 7.42 (1H, s), 7.48 (2H, d, J=7.9 Hz),7.97 (2H, d, J=7.9 Hz), 8.40 (1H, s).

Step 2(1R,3S,4R)—N¹-{[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}-4-methoxy-N³-methyl-N³-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

A mixture of the compound (0.454 g) obtained in the above Step 1, 1Nhydrochloric acid (0.739 mL) and ethanol (3 mL) was stirred at roomtemperature for 5 min, and concentrated. Acetonitrile (10 mL) was addedto the obtained residue, and the precipitated solid was collected byfiltration, and dried to give the title compound (0.413 g) as a solid.

¹H-NMR (DMSO-D₆) δ: 1.88-1.95 (1H, m), 2.32-2.39 (2H, m), 2.46-2.49 (1H,m), 3.17 (3H, s), 3.41 (3H, s), 3.55-3.67 (1H, m), 3.99-4.04 (1H, m),4.01 (3H, s), 4.07 (3H, s), 4.09 (2H, q, J=11.0 Hz), 4.25-4.30 (2H, m),5.07-5.15 (1H, m), 7.67 (1H, s), 7.71 (2H, d, J=8.6 Hz), 7.77 (1H, s),8.20 (2H, d, J=8.6 Hz), 8.39 (1H, s), 9.29 (2H, br s).

MS (m/z): 589 (M+H)⁺.

Example 128(1R,2S,4R)-4-({[(1s,4S)-4-(5,6-dimethoxypyridazin-3-yl)cyclohexyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride(1R,2S,4R)-4-({[(1r,4R)-4-(5,6-dimethoxypyridazin-3-yl)cyclohexyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olhydrochloride

The compound (0.220 g) obtained in Step 2 of Reference Example C-15 wassuspended in dichloromethane (5.3 mL), and DIPEA (0.371 mL) was addedthereto. The mixture was stirred at room temperature for 1 hr. Thecompound (0.140 g) obtained in Step 2 of Reference Example D-78 andacetic acid (0.213 mL) were added thereto, and the mixture was stirredfor 15 min. Sodium triacetoxyborohydride (0.384 g) was added to themixture, and the mixture was stirred overnight at room temperature.Saturated aqueous sodium bicarbonate solution was added to the reactionsolution, and the mixture was subjected to extraction operation. Theorganic layer was dried over anhydrous sodium sulfate, and the solventwas evaporated under reduced pressure. The residue was purified bysilica gel chromatography (dichloromethane/methanol) to give a solid.The solid was purified by high-performance liquid chromatography(CHIRALPAK (registered trademark, Daicel Corporation) IG, mobile phase:n-hexane/ethanol) to give a free form of 128A (an earlier elutedcomponent, 0.026 g) and a free form of 128B (a later eluted component0.100 g), respectively as a solid.

the free form of 128A

¹H-NMR (CDCl₃) δ: 1.68-2.08 (12H, m), 2.26-2.35 (1H, m), 2.68-2.74 (2H,m), 2.85-2.92 (1H, m), 3.35-3.42 (1H, m), 3.50 (3H, s), 3.54 (1H, d,J=10.5 Hz), 3.59 (1H, d, J=10.5 Hz), 3.93 (3H, s), 3.96 (3H, s), 4.16(3H, s), 4.43 (1H, t, J=4.0 Hz), 5.12 (1H, td, J=10.0, 5.0 Hz), 6.61(1H, s), 7.32 (1H, s).

MS (m/z): 611 (M+H)⁺.

the free form of 128B

¹H-NMR (CDCl₃) δ: 1.10-1.23 (2H, m), 1.57-2.09 (10H, m), 2.27-2.36 (1H,m), 2.57 (2H, d, J=7.0 Hz), 2.76 (1H, t, J=12.5 Hz), 3.32-3.38 (1H, m),3.52 (3H, s), 3.54 (1H, d, J=10.5 Hz), 3.59 (1H, d, J=10.5 Hz), 3.92(3H, s), 3.96 (3H, s), 4.15 (3H, s), 4.42-4.46 (1H, m), 5.10-5.18 (1H,m), 6.59 (1H, s), 7.32 (1H, s).

MS (m/z): 611 (M+H)⁺.

The free form of 128A (25 mg) obtained in the above step was dissolvedin ethanol (0.5 mL), 1N hydrochloric acid ethanol solution (41 μl) wasadded thereto, and the mixture was concentrated under reduced pressure.Ethyl acetate (1 mL) was added to the residue, and the precipitatedsolid was collected by filtration to give one compound (128A) (28 mg) ofthe title compounds, as a solid.

¹H-NMR (DMSO-D₆) δ: 1.60-2.12 (11H, m), 2.23-2.42 (2H, m), 2.85-3.10(3H, m), 3.40 (3H, s), 3.53-3.64 (1H, m), 3.84 (3H, s), 3.92 (3H, s),3.96-4.02 (5H, m), 4.34-4.40 (1H, m), 4.77-4.85 (1H, m), 5.14-5.20 (1H,m), 7.19 (1H, br s), 7.63 (1H, s), 8.87-8.60 (2H, m).

MS (m/z): 611 (M+H)⁺.

Similarly, the free form of 128B (95.0 mg) was dissolved in ethanol (2mL), 1N hydrochloric acid ethanol solution (0.160 mL) was added thereto,and the mixture was concentrated under reduced pressure. Ethyl acetate(2 mL) was added to the residue, and the precipitated solid wascollected by filtration to give the other compound (128B) (0.103 g) ofthe title compounds, as a solid.

¹H-NMR (DMSO-D₆) δ: 1.10-1.24 (2H, m), 1.54-2.07 (9H, m), 2.22-2.44 (2H,m), 2.69-2.92 (3H, m), 3.40 (3H, s), 3.52-3.63 (1H, m), 3.85 (3H, s),3.89 (3H, s), 3.94-4.04 (5H, m), 4.33-4.40 (1H, m), 4.77-4.87 (1H, m),5.11-5.19 (1H, m), 7.09 (1H, br s), 7.62 (1H, s), 8.98-8.67 (2H, m).

MS (m/z): 611 (M+H)⁺.

separation condition (analysis) CHIRALPAK (registered trademark, DaicelCorporation) IG, size 0.46 cm×25 cm, flow rate 1.0 mL/min, mobile phasen-hexane/ethanol=20/80, temperature 40° C.

the free form of 128A retention time 8.5 min, the free form of 128Bretention time 11.7 min

separation condition (preparative) CHIRALPAK (registered trademark,Daicel Corporation) IG, size 2 cm×25 cm, flow rate 15.0 mL/min, mobilephase n-hexane/ethanol=20/80, temperature 40° C.

the free form of 128A retention time 9.9 min, the free form of 128Bretention time 12.2 min

Example 129(1R,2S,4R)-4-({[4-(5-methoxypyridin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olhydrochloride

The title compound was obtained in the same manner as in Step 1 and Step2 of Example 27, using the compound obtained in Step 2 of ReferenceExample C-14 and the compound obtained in Reference Example D-65.

¹H-NMR (DMSO-D₆) δ: 1.83-1.90 (1H, m), 2.28-2.34 (1H, m), 2.44-2.54 (2H,m), 3.44 (3H, s), 3.55-3.64 (1H, m), 3.93 (3H, s), 4.09 (2H, q, J=10.9Hz), 4.23-4.28 (2H, m), 4.33-4.37 (1H, m), 4.91-4.99 (1H, m), 5.20 (1H,d, J=4.3 Hz), 7.67-7.69 (1H, m), 7.72-7.76 (3H, m), 7.86 (2H, d, J=7.9Hz), 8.32 (1H, d, J=2.4 Hz), 8.36 (1H, s), 8.54 (1H, d, J=1.2 Hz),9.52-9.64 (2H, m).

Example 130(1R,3S)—N³-{[4-(2-methoxy-1,3-thiazol-5-yl)phenyl]methyl}-N¹-methyl-N¹-[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]cyclopentane-1,3-diaminehydrochloride

The title compound was obtained in the same manner as in Step 2 and Step3 of Example 11, using the compound obtained in Step 1 of Example 11 and2-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole.

¹H-NMR (DMSO-D₆) δ: 1.86-1.88 (1H, m), 1.95-2.09 (4H, m), 2.31-2.38 (1H,m), 3.29 (3H, s), 3.56-3.63 (1H, br m), 4.06-4.15 (7H, m), 5.24-5.29(1H, br m), 7.51-7.54 (3H, m), 7.58 (2H, d, J=8.5 Hz), 7.74 (1H, s),8.40 (1H, s), 9.35 (1H, br s), 9.48 (1H, br s), 11.57 (1H, d, J=2.4 Hz).

MS (m/z): 534 (M+H)⁺.

Example 131(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olsuccinate

(1R,2S,4R)-4-({[4-(5,6-Dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol(25.8 g) obtained in the same manner as in Step 1 of Example 25 wassuspended in 2-propanol (103 mL), and water (12.9 mL) was added thereto.Succinic acid (5.56 g) was added to the mixture, water (12.9 mL) wasadded thereto, and the mixture was stirred at room temperature for 4 hr.The insoluble substance was collected by filtration while washing with2-propanol (150 mL), and dried to give the title compound (30.1 g) ascrystals.

¹H-NMR (DMSO-D₆) δ: 1.50-1.58 (1H, m), 2.04-2.19 (2H, m), 2.27-2.35 (1H,m), 2.37 (4H, s), 3.14-3.24 (1H, m), 3.42 (3H, s), 3.88-3.94 (2H, m),3.99 (3H, s), 4.05-4.11 (5H, m), 4.28-4.33 (1H, m), 4.80-4.90 (1H, m),7.54 (2H, d, J=8.5 Hz), 7.61 (1H, s), 7.72 (1H, s), 8.08 (2H, d, J=8.5Hz), 8.33 (1H, s).

MS (m/z): 575 (M+H)⁺.

[α]_(D) ²⁰ −28.3 (c 1.00, DMSO)

elemental analysis found value. C, 53.62; H, 5.11; F, 8.27; N, 12.05; S,4.58.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 1.

Table 3 shows peaks with a relative intensity of 35 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 1 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 3 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 4.66 18.95 48 6 18.68 4.75 100 2 7.02 12.58 87 7 21.344.16 94 3 14.10 6.28 73 8 24.52 3.63 35 4 16.68 5.31 45 9 25.54 3.48 385 17.46 5.08 58 10 28.22 3.16 46

Example 132(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olbenzenesulfonate

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol(200.31 mg) obtained in the same manner as in Step 1 of Example 25 andbenzenesulfonic acid (57.89 mg) was added 80% hydrous acetone (1639 μL),and the mixture was stirred at room temperature about 24 hr. Theprecipitated solid was collected, and dried overnight at roomtemperature to give the title compound (220.45 mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.72-1.79 (1H, m), 2.26-2.47 (3H, m), 3.44 (3H, s),3.58-3.68 (1H, m), 4.01 (3H, s), 4.04-4.14 (5H. m), 4.26-4.42 (3H, m),4.90-4.98 (1H, m), 5.22 (1H, d, J=4.0 Hz), 7.28-7.34 (3H, m), 7.57-7.61(2H, m), 7.67 (1H, s), 7.69 (2H, d, J=8.5 Hz), 7.75 (1H, s), 8.20 (2H,d, J=8.5 Hz), 8.36 (1H, s), 8.98-9.17 (2H, m).

MS (m/z): 575 (M+H)⁺.

elemental analysis found value. C, 50.24; H, 5.06; F, 7.64; N, 10.57; S,8.21.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 2.

Table 4 shows peaks with a relative intensity of 24 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 2 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 4 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 10.92 8.10 30 6 18.16 4.88 22 2 11.70 7.56 27 7 22.184.00 30 3 12.40 7.13 100 8 22.62 3.93 35 4 15.00 5.90 35 9 23.86 3.73 265 17.38 5.10 24 10 24.20 3.67 24

Example 133(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olmaleate

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol(300.14 mg) obtained in the same manner as in Step 1 of Example 25 andmaleic acid (63.66 mg) was added 80% hydrous 2-propanol (5455 μL), andthe mixture was stirred at 40° C. for about 24 hr. The precipitatedsolid was collected, and dried overnight at room temperature to give thetitle compound (304.38 mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.71-1.79 (1H, m), 2.25-2.48 (3H, m), 3.45 (3H, s),3.57-3.67 (1H, m), 4.01 (3H, s), 4.04-4.14 (5H, m), 4.25-4.41 (3H, m),4.90-4.98 (1H, m), 5.15-5.27 (1H, m), 6.02 (2H, s), 7.68 (1H, s), 7.68(2H, d, J=8.0 Hz), 7.75 (1H, s), 8.20 (2H, d, J=8.0 Hz), 8.37 (1H, s),9.05 (2H, br s).

MS (m/z): 575 (M+H)⁺.

elemental analysis found value. C, 53.61; H, 4.81; F, 8.25; N, 12.15; S,4.67.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 3.

Table 5 shows peaks with a relative intensity of 42 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 3 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 5 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 4.64 19.03 47 6 16.76 5.29 74 2 7.02 12.58 43 7 18.544.78 83 3 7.46 11.84 59 8 19.76 4.49 60 4 11.14 7.94 56 9 21.26 4.18 425 14.04 6.30 61 10 22.62 3.93 100

Example 134(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olfumarate

To the compound (200.38 mg) obtained in the same manner as in Step 1 ofExample 25 and fumaric acid (42.50 mg) was added 80% hydrous 2-propanol(1638 μL), and the mixture was stirred at 40° C. for about 24 hr. Theprecipitated solid was collected, and dried overnight at roomtemperature to give the title compound (236.21 mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.57-1.66 (1H, m), 2.14-2.22 (2H, m), 2.30-2.39 (1H,m), 3.21-3.31 (1H, m), 3.42 (3H, s), 3.97-4.01 (5H, m), 4.03-4.13 (5H,m), 4.28-4.33 (1H, m), 4.82-4.91 (1H, m), 6.55 (2H, s), 7.58 (2H, d,J=8.0 Hz), 7.62 (1H, s), 7.73 (1H, s), 8.10 (2H, d, J=8.0 Hz), 8.33 (1H,s).

MS (m/z):575 (M+H)⁺.

elemental analysis found value. C, 48.52; H, 5.31; F, 7.63; N, 10.89; S,4.27.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 4.

Table 6 shows peaks with a relative intensity of 51 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 4 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 6 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 4.80 18.39 73 6 17.62 5.03 100 2 7.94 11.13 92 7 18.144.89 71 3 9.66 9.15 99 8 20.46 4.34 59 4 11.56 7.65 67 9 21.36 4.16 53 514.56 6.08 73 10 24.46 3.64 51

Example 135 crystal of(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol(20.17 mg) obtained in the same manner as in Step 1 of Example 25 wereadded 2-propanol (81 μL) and water (323 μL), and the mixture was stirredat room temperature for about 24 hr. The obtained crystals werecollected, and dried overnight at room temperature to give the titlecompound (13.89 mg).

elemental analysis found value. C, 53.01; H, 5.26; F, 8.97; N, 12.49; S,4.84.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 5.

Table 7 shows peaks with a relative intensity of 12 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 5 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 7 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 7.14 12.37 100 6 23.40 3.80 15 2 8.76 10.09 14 7 24.403.65 12 3 12.26 7.21 38 8 24.86 3.58 12 4 14.30 6.19 25 9 25.34 3.51 165 17.52 5.06 16 10 25.90 3.44 13

Example 136(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olfumarate

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol(20.76 mg) obtained in the same manner as in Step 1 of Example 25 andfumaric acid (4.150 mg) was added 80% hydrous 2-propanol (415 μL), andthe mixture was stirred at room temperature for about 24 hr. Theprecipitated solid was collected, and dried overnight at roomtemperature to give the title compound (22.13 mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.55-1.63 (1H, m), 2.08-2.19 (2H, m), 2.28-2.37 (1H,m), 3.18-3.28 (1H, m), 3.39 (3H, s), 3.82 (3H, s), 3.92-4.03 (7H, m),4.06 (3H, s), 4.29-4.34 (1H, m), 4.71-4.80 (1H, m), 6.55 (2H, s), 7.57(2H, d, J=8.0 Hz), 7.61 (1H, br s), 7.62 (1H, s), 8.09 (2H, d, J=8.0Hz). elemental analysis found value. C, 50.58; H, 4.91; F, 7.58; N,10.89; S, 4.15.

MS (m/z): 605 (M+H)⁺.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 6.

Table 8 shows peaks with a relative intensity of 20 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 6 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 8 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 8.06 10.96 100 6 18.56 4.78 22 2 12.22 7.24 58 7 20.084.42 22 3 12.52 7.06 39 8 23.48 3.79 23 4 15.14 5.85 20 9 24.28 3.66 245 17.54 5.05 23 10 25.00 3.56 23

Example 137(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olmucate

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol(20.10 mg) obtained in the same manner as in Step 1 of Example 25 andmucic acid (7.421 mg) was added 80% hydrous 2-propanol (402 μL), and themixture was stirred at room temperature for about 24 hr. Theprecipitated solid was collected, and dried overnight at roomtemperature to give the title compound (16.85 mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.53-1.61 (1H, m), 2.06-2.21 (2H, m), 2.28-2.37 (1H,m), 3.17-3.27 (1H, m), 3.39 (3H, s), 3.72 (2H, s), 3.82 (3H, s),3.92-4.03 (7H, m), 4.06 (3H, s), 4.14 (2H, s), 4.30-4.35 (1H, m),4.71-4.81 (1H, m), 7.56 (2H, d, J=8.0 Hz), 7.61 (1H, s), 7.61 (1H, brs), 8.09 (2H, d, J=8.0 Hz).

MS (m/z): 605 (M+H)⁺. elemental analysis found value. C, 46.77; H, 5.19;F, 6.71; N, 9.62; S, 3.61.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 7.

Table 9 shows peaks with a relative intensity of 22 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 7 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 9 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 6.56 13.46 54 6 18.86 4.70 22 2 9.44 9.36 48 7 19.604.53 40 3 9.94 8.89 59 8 22.68 3.92 27 4 13.20 6.70 100 9 25.10 3.54 305 18.22 4.87 40 10 28.70 3.11 23

(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-oladipate

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol(19.68 mg) obtained in the same manner as in Step 1 of Example 25 andadipic acid (5.019 mg) was added 80% hydrous 2-propanol (394 μL), andthe mixture was stirred at room temperature for about 24 hr. Theprecipitated solid was collected, and dried overnight at roomtemperature to give the title compound (19.75 mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.44-1.53 (5H, m), 1.93-2.31 (7H, m), 3.01-3.10 (1H,m), 3.38 (3H, s), 3.76-3.86 (5H, m), 3.93-4.03 (5H, m), 4.05 (3H, s),4.27-4.32 (1H, m), 4.68-4.78 (1H, m), 7.50 (2H, d, J=8.5 Hz), 7.59 (2H,br s), 8.05 (2H, d, J=8.5 Hz).

MS (m/z): 605 (M+H)⁺.

elemental analysis found value. C, 50.67; H, 5.93; F, 7.28; N, 10.33; S,3.98.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 8.

Table 10 shows peaks with a relative intensity of 10 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 8 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 10 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 5.88 15.02 100 6 13.70 6.46 48 2 6.20 14.24 30 7 15.665.65 22 3 9.18 9.63 11 8 17.82 4.97 10 4 10.34 8.55 15 9 18.48 4.80 13 512.50 7.08 11 10 22.16 4.01 17

Example 139(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-olsuccinate

To(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{[2-methoxy-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl](methyl)amino}cyclopentan-1-ol(20.27 mg) obtained in the same manner as in Step 1 of Example 25 wasadded 80% hydrous 2-propanol (368 μL), 1 mol/L aqueous succinic acidsolution (81 μL) was added thereto, and the mixture was stirred at roomtemperature for about 24 hr. The precipitated solid was collected, anddried overnight at room temperature to give the title compound (14.41mg) as crystals.

¹H-NMR (DMSO-D₆) δ: 1.50-1.58 (1H, m), 2.02-2.19 (2H, m), 2.26-2.34 (1H,m), 2.37 (4H, s), 3.13-3.23 (1H, m), 3.39 (3H, s), 3.82 (3H, s),3.86-4.03 (7H, m), 4.06 (3H, s), 4.29-4.34 (1H, m), 4.70-4.80 (1H, m),7.54 (2H, d, J=8.0 Hz), 7.61 (2H, br s), 8.08 (2H, d, J=8.0 Hz).

MS (m/z): 605 (M+H)⁺.

[α]_(D) ²⁰ −43.6 (c 1.00, DMSO)

elemental analysis found value. C, 49.98; H, 5.30; F, 7.33; N, 10.73; S,4.14.

The powder X-RAY diffraction of the obtained crystal is shown in FIG. 9.

Table 11 shows peaks with a relative intensity of 26 or more when themaximum peak intensity is 100 in the diffraction pattern FIG. 9 of thepowder X-RAY diffraction (CuKα, λ=1.54 angstroms, scanningspeed=20°/min).

TABLE 11 peak relative peak relative No. 2θ d value intensity No. 2θ dvalue intensity 1 4.60 19.19 39 6 11.16 7.92 31 2 6.60 13.38 100 7 12.007.37 52 3 7.74 11.41 90 8 12.44 7.11 40 4 8.02 11.01 100 9 13.22 6.69 305 9.26 9.54 26 10 19.66 4.51 41

Formulation Example Formulation Example 1 (Injection)

1.5% by weight of the Example compound is stirred in 10% by volume ofpropylene glycol, then the volume is adjusted to a predetermined valuewith water for injection, and the mixture is sterilized to give aninjection.

Formulation Example 2 (Hard Capsule)

100 mg of the powdered Example compound, 128.7 mg of lactose, 70 mg ofcellulose and 1.3 mg of magnesium stearate are mixed and passed througha 60 mesh sieve, and the resulting powder is put in 250 mg No. 3 gelatincapsules to give capsules.

Formulation Example 3 (Tablet)

100 mg of the powdered Example compound, 124 mg of lactose, 25 mg ofcellulose and 1 mg of magnesium stearate are mixed and tableted with atableting machine to give 250 mg of tablets per tablet. This tablet canbe sugar-coated as needed.

Experimental Example

The pharmacological activity of the compound of the present inventionwas confirmed by the following tests.

[Experimental Example 1] Evaluation of Binding Inhibitory ActivityBetween Menin and MLL

10 μL of the reaction solution (50 mM Tris-HCl(pH7.5), 50 mM NaCl, 0.01%Triton X-100, 0.01% Bovine serum albumin, 3 mM TCEP) containing any ofthe compound of Examples 1 to 130, 1 nM menin (Flag tagged, DaiichiSankyo RD Novare Co., Ltd.) and 10 nM biotinylated MLL1 peptide (1-46aa:Scrum Co., Ltd.) was added to a 384-well plate, and reacted at roomtemperature for minutes. Then, 10 μL of a mixed solution (10 μg/mL each)of Anti-FLAG AlphaLISA Acceptor beads (PerkinElmer Co., Ltd., AL112C)and Streptavidin-coated AlphaScreen Donor beads (PerkinElmer Co., Ltd.,6760002) was added thereto, and the mixture was reacted at roomtemperature for 1 hr. Then, the fluorescence signal by AlphaLISA(registered trademark, PerkinElmer Co., Ltd.) was measured using a platereader (PerkinElmer Co., Ltd., EnVisionXcite). Based on the measuredsignals, the binding inhibitory rates of the compounds of Examples 1 to130 at each concentration were calculated, and the obtained data wasanalyzed with the medical statistical analysis software GraphPad Prism(GraphPad Software, Inc.) to obtain the IC₅₀ values.

[Experimental Example 2] Evaluation of Cell Growth Inhibitory Activity

RPMI1640 medium supplemented with 10% FBS was used as a culture mediumfor each cells (MV-4-11, MOLM-13 cells, human AML cells, K562 cells,human CML cells). Each cell was purchased from American Type CultureCollection (ATCC). After diluting and preparing each drug (Examplecompound) with Freedom EVO 150 (Tecan Trading AG) (common ratio of 2, 10concentrations from 10 mM or 5 mM to 20 μM or 10 μM), the drug wasseeded at 40 nL/well using Echo555 (Labcyte Inc.) on a 384-well tissueculture plate (#3712, Corning Inc.) (final concentration 10 μM or 5 μMto 20 nM or 10 nM). The prepared drug-containing plate was stored at−30° C. until use, and thawed before use.

The suspension of each cells was prepared with 10% FBS RPMI1640 culturemedium so as to be 1000 cells/mL (K562) or 10000 cells/mL (MV-4-11,MOLM-13), and seeded on the drug-containing plate (40 μL/well) (day 0).The cells were cultured for additional 3 or 7 days. ATP measurementreagent (CellTiter-Glo (registered trademark, Promega Corporation) 2.0Assay, model number G9242, Promega Corporation) was added to each wellat 10 μL/well, on the day of the drug addition (day 0) and 3 days (day3) or 7 days (day 7) after the drug addition. After affixing a blacksticker on the bottom surface, the mixture was stirred using amicroplate mixing deaeration machine (model name, Weltornado FK-62,Sakaki Dengyo Co., Ltd.) (stirring conditions, revolution 9, rotation 7,time 12). The luminescence (cps) was measured using a luminescencedetector of a microplate reader (model name EnVision 2102-0010,PerkinElmer Co., Ltd.) (N=4).

As an index of the cell growth inhibitory activity, the concentration(GI₅) that suppresses cell growth by 50% was calculated using EXCEL2010(Microsoft Corporation). The cell growth of the drug-added group fromday 0 to day 3 or day 7 was calculated as T/C %, when the cell growth ofthe drug-free group was considered as 100%. GI₅₀ was calculated byGROWTH function (exponential regression) using two concentrationssandwiching the concentration (T/C %=50%) that suppresses cell growth by50%, and T/C %.

The results in Experimental Examples 1 and 2 are shown in Table 12-1 toTable 12-4.

TABLE 12-1 Ex. cell free GI₅₀ (μM) Day3 GI₅₀ (μM) Day7 No. IC₅₀ (nM)K562 MV4;11 MOLM-13 K562 MV4;11 MOLM-13  1 493.7 5.09 1.79 NT NT NT NT 2 12.8 2.81 0.330 NT NT NT NT  3 81.8 3.78 0.748 NT NT NT NT  4 172.44.37 1.32 NT NT NT NT  5 855.5 >10.0 3.12 NT NT NT NT  6 81.9 >10.00.679 NT 6.89 0.439 1.10  7 150.7 1.67 1.36 1.69 0.95 0.908 1.82  8 8.82.73 0.408 0.87 1.44 0.201 0.592  9 22.2 2.95 0.585 1.61 NT NT NT 10199.0 >10.0 >10.0 >10.0 NT NT NT 11 63.9 6.66 1.30 3.28 3.82 0.711 1.1912 33.8 6.72 0.702 2.72 NT NT NT 13 7.6 2.16 0.273 1.01 NT NT NT 14 27.42.23 0.655 1.48 NT NT NT 15A 21.6 >10.0 0.269 4.41 NT NT NT 15B 402.85.42 1.88 3.15 NT NT NT 16 1868.0 7.43 5.18 6.04 NT NT NT 17A 2262.08.66 2.25 3.29 5.27 1.58 3.35 17B 13.3 >10.0 0.170 5.00 7.25 0.118 0.31018A 10.3 5.70 0.260 2.83 5.01 0.131 0.448 18B 182.2 5.18 2.16 3.08 4.711.47 3.97 19A 33.5 5.51 0.489 3.20 5.39 0.295 0.906 19B 1814.0 5.45 3.184.03 5.41 2.60 5.75 20A 3.9 5.73 0.367 2.90 2.86 0.177 0.352 20B 16.49.03 0.331 4.02 4.65 0.226 0.379 21 7.0 5.75 0.304 4.50 9.60 0.173 0.50922 3.8 1.37 0.189 0.675 0.925 0.0971 0.262 23 2.8 1.55 0.106 0.594 0.5860.0584 0.155 24 4.1 3.23 0.0609 0.873 2.57 0.0302 0.104 25 1.0 >5.000.0253 0.224 3.47 0.0180 0.0400 26 3.8 2.74 0.121 1.39 1.93 0.0715 0.249

TABLE 12-2 27 1.3 3.37 0.0152 0.139 2.10 0.0123 0.0313 28 1163.0 6.315.34 NT NT NT NT 29 463.6 >10.0 1.26 NT 7.77 1.07 1.66 30 44.6 2.340.867 NT NT NT NT 31 46.4 >10.0 1.29 NT NT NT NT 32 94.9 5.59 2.43 NT NTNT NT 33 2419.0 6.44 5.22 4.82 NT NT NT 34 412.7 4.49 0.420 0.718 2.770.106 0.444 35 1222.0 6.97 4.34 5.72 NT NT NT 36 636.0 >10.0 3.13 3.14NT NT NT 37 700.8 >10.0 5.21 7.21 NT NT NT 38 1850.0 >10.0 6.09 >10.0 NTNT NT 39 2067.0 >10.0 8.14 >10.0 NT NT NT 40 265.5 5.53 2.62 4.54 4.162.16 4.33 41 471.6 2.67 2.46 2.54 NT NT NT 42 104.8 5.09 1.43 3.11 2.871.14 1.96 43 76.2 3.02 1.18 2.41 1.72 0.877 1.74 44 335.2 2.75 1.89 2.731.91 1.53 2.93 45 33.3 3.27 0.865 1.78 2.00 0.701 1.40 46 39.4 6.420.490 2.69 NT NT NT 47 97.2 6.20 0.704 2.22 NT NT NT 48 1417.0 3.45 1.772.21 NT NT NT 49 24.0 9.74 0.598 2.52 3.18 0.490 1.13 50 9.7 5.69 0.2002.53 3.29 0.0812 0.330 51 10.1 3.20 0.137 1.24 2.06 0.0621 0.259 52612.0 5.93 2.39 3.37 NT NT NT 53 17.7 NT NT NT 0.908 0.395 0.826 54 1.03.13 0.0669 1.33 2.18 0.0279 0.0579 55 9.8 1.90 0.149 0.614 1.32 0.08950.221 56 43.6 1.26 0.769 1.02 0.809 0.468 1.07 57 61.2 6.30 1.16 2.143.03 0.689 1.56 58 15.2 1.03 0.399 0.804 0.493 0.230 0.485 59 33.0 1.750.894 1.22 1.05 0.451 1.13 60 2.4 3.40 0.0776 0.614 2.39 0.0325 0.080761 1.9 6.25 0.110 0.974 5.74 0.0472 0.129

TABLE 12-3 62 2.0 2.69 0.113 1.06 2.59 0.0666 0.199 63 19.9 2.55 0.6821.88 1.48 0.445 1.21 64 2.2 2.18 0.0563 0.358 1.67 0.0355 0.0933 65 2.52.85 0.0882 1.46 2.64 0.0557 0.143 66 6.7 1.61 0.286 0.978 0.865 0.1770.540 67 2.1 4.00 0.0676 1.42 2.36 0.0440 0.130 68 1.7 4.93 0.0462 1.932.85 0.0207 0.0882 69 2.8 1.53 0.0382 0.630 1.41 0.0233 0.0637 70 3.82.57 0.0673 0.948 2.53 0.0351 0.111 71 0.6 2.80 0.285 0.964 1.77 0.1570.469 72 10.7 NT NT NT 1.00 0.230 0.519 73 6.5 NT NT NT 0.723 <0.01950.0389 74 6.5 NT NT NT 2.55 0.0455 0.122 75 17.2 NT NT NT 1.47 0.05710.194 76 8.6 4.80 0.183 1.70 2.70 0.0963 0.268 77 10.4 2.60 0.350 1.171.62 0.160 0.501 78 8.5 3.53 0.219 1.72 1.99 0.114 0.378 79 3.5 4.340.313 1.64 2.12 0.173 0.482 80 15.4 2.79 0.434 1.88 2.16 0.246 0.705 816.1 2.22 0.0544 0.569 1.61 0.0246 0.0943 82 22.8 1.41 0.112 0.521 1.330.0505 0.177 83 4.9 2.38 0.345 1.45 1.52 0.179 0.646 84 2.7 5.28 0.02350.652 3.39 <0.0195 0.0455 85 4.0 2.04 0.137 0.742 0.921 0.0749 0.240 8620.9 3.97 0.608 1.78 1.96 0.372 0.949 87 6.6 1.76 0.165 0.803 1.23 0.1140.423 88 27.8 2.01 0.446 1.48 1.76 0.327 1.01 89 5.6 >10.0 0.07617.18 >10.0 0.0467 0.185 90 1.2 0.750 <0.0195 0.324 0.636 <0.0195 0.036491 1.3 0.769 <0.0195 0.206 0.542 <0.0195 0.0526 92 1.5 3.48 0.0891 0.8262.79 0.0493 0.120 93 1.6 1.38 0.0552 0.659 1.41 0.0357 0.107 94 2.1 5.040.143 1.54 2.82 0.0751 0.227 95 1.2 5.08 0.0454 1.10 1.78 0.0239 0.066896 2.7 3.04 0.126 0.268 2.61 0.158 0.281

TABLE 12-4  97 3.9 3.21 0.295 0.852 2.46 0.187 0.285  98 1.9 5.57 0.1090.411 2.10 0.0628 0.105  99 1.1 3.80 0.0626 0.187 1.84 0.0365 0.0562 1002.5 >10.0 0.0236 0.137 5.94 <0.0195 0.0598 101 2.7 2.76 0.0563 0.2162.03 0.0502 0.132 102 2.7 >10.0 0.0393 0.258 >10.0 0.0271 0.0742 1031.4 >10.0 0.028 0.13 8.00 0.032 0.073 104 6.3 >5.00 0.097 0.70 >5.000.066 0.187 105 8.0 >5.00 0.156 1.63 3.37 0.129 0.285 106 0.7 NT NT NT0.435 0.0248 0.0597 107 1.3 NT NT NT 4.91 0.0270 0.0647 108 1.3 NT NTNT >5.00 0.0234 0.0622 109 0.7 NT NT NT >5.00 0.0133 0.0312 110 2.1 NTNT NT 1.60 0.0763 0.253 111 42.3 NT NT NT 1.20 0.634 0.751 112 0.8 NT NTNT 2.56 0.0265 0.995 113 29.2 NT NT NT 1.66 0.0997 1.15 114 1.5 NT NT NT1.94 0.0440 0.143 115 10.9 NT NT NT >5.00 0.222 0.974 116 1.0 NT NT NT1.00 0.0133 0.0592 117 1.4 NT NT NT 1.56 0.0468 0.184 118 1.0 NT NT NT2.70 0.0254 0.127 119 0.7 NT NT NT 0.859 0.0157 0.0637 120 5.8 NT NT NT2.16 0.225 0.729 121 0.8 NT NT NT 0.894 0.0381 0.0495 122 1.7 NT NT NT2.27 0.232 0.236 123 0.6 NT NT NT 1.69 0.0223 0.0284 124 182.1 NT NT NT4.82 0.624 0.922 125 1.5 NT NT NT 3.75 0.0769 0.154 126 7.5 >10.0 0.2575.03 8.84 0.137 0.396 127 1.6 7.14 0.169 2.15 4.26 0.092 0.242 128A 18.4NT NT NT >5.00 0.728 1.451 128B 6.6 NT NT NT >5.00 0.201 0.501 129 5.55.21 0.0968 1.25 2.74 0.0378 0.104 130 6.5 1.90 0.241 0.894 NT NT NT

NT: Not Tested. [Experimental Example 3-1] Evaluation of AntitumorActivity in MV-4-11 Cell Subcutaneous Transplant Model

MV-4-11 cells were transplanted subcutaneously to the right abdomen offemale FOX CHASE SCIDC.B.17/Icr-scid/scidJcl mice at a ratio of 1×10⁷cells/head, and after 17 days, the mice were grouped into 6 animals pereach group, based on the estimated tumor volume (major axis×minoraxis×minor axis/2). MV-4-11 cells were purchased from ATCC. The femaleFOX CHASE SCID C.B.17/Icr-scid/scidJcl mice were purchased from ClaireJapan. From the day after the grouping, the compound of Example 26 wasorally administered on a schedule of once daily for 17 days everyday(qd×17), at a dose setting of 25, 50 or 100 mg/kg/day. The compound wasadministered as a suspension in 0.5% methylcellulose (MC). For thecompound-free group, 0.5% MC was administered as a solvent. Theestimated individual tumor volume was measured from the day of thegrouping to 34 days after the transplantation (test end date).

[Experimental Example 3-2] Evaluation of Antitumor Activity in MV-4-11Cell Subcutaneous Transplant Model

MV-4-11 cells were transplanted subcutaneously to the right abdomen offemale FOX CHASE SCID C.B.17/Icr-scid/scidJcl mice at a ratio of 1×107cells/head, and after 17 days, the mice were grouped into 6 animals pereach group, based on the estimated tumor volume (major axis x minor axisx minor axis/2). MV-4-11 cells were purchased from ATCC. The female FOXCHASE SCID C.B.17/Icr-scid/scidJcl mice were purchased from ClaireJapan. From the day of the grouping, the compound of Example 25 or 27was orally administered on a schedule of once daily for 18 days everyday(qd×18), at a dose setting of 12.5, 25, 50 or 100 mg/kg/day. Thecompound was administered as a suspension in 0.5% MC. For thecompound-free group, 0.5% MC was administered as a solvent. Theestimated individual tumor volume was measured from the day of thegrouping to 34 days after the transplantation (test end date).

[Experimental Example 3-3] MV-4-Llcell Evaluation of Antitumor Activityin MV-4-11 Cell Subcutaneous Transplant Model

MV-4-11 cells were transplanted subcutaneously to the right abdomen offemale FOX CHASE SCID C.B.17/Icr-scid/scidJcl mice at a ratio of 1×107cells/head, and after 14 days, the mice were grouped into 6 animals pereach group, based on the estimated tumor volume (major axis×minoraxis×minor axis/2). MV-4-11 cells were purchased from ATCC. The femaleFOX CHASE SCID C.B.17/Icr-scid/scidJcl mice were purchased from ClaireJapan. From the day after the grouping, the compound of Example 68, 25or 27 was orally administered on a schedule of once daily for 17 dayseveryday (qd×17), at a dose setting of 25, 50 or 100 mg/kg/day. Thecompound was administered as a suspension in 0.5% MC. For thecompound-free group, 0.5% MC was administered as a solvent. Theestimated individual tumor volume was measured from the day of thegrouping to 31 days after the transplantation (test end date).

[Experimental Example 3-4] Evaluation of Antitumor Activity in MV-4-11Cell Subcutaneous Transplant Model

MV-4-11 cells were transplanted subcutaneously to the right abdomen offemale FOX CHASE SCID C.B.17/Icr-scid/scidJcl mice at a ratio of 1×107cells/head, and after 15 days, the mice were grouped into 6 animals pereach group, based on the estimated tumor volume (major axis x minor axisx minor axis/2). MV-4-11 cells were purchased from ATCC. The female FOXCHASE SCID C.B.17/Icr-scid/scidJcl mice were purchased from ClaireJapan. From the day after the grouping, the compound of Example 60 or 67was orally administered on a schedule of once daily for 16 days everyday(qd×16), at a dose setting of 25, 50 or 100 mg/kg/day. The compound wasadministered as a suspension in 0.5% MC. For the compound-free group,0.5% MC was administered as a solvent. The estimated individual tumorvolume was measured from the day of the grouping to 32 days after thetransplantation (test end date).

The antitumor activities in MV4:11 cell subcutaneous transplant model inExperimental Examples 3-1 to 3-4 were calculated by the followingformula on the end date of each test.

Tumor growth inhibitory rate %=(1−TVCt/TVCc)×100 TVC=(Individual tumorvolume on the test end date)−(Individual tumor volume on the groupingdate)

TVCt: TVC mean value of the compound administration groupTVCc: TVC mean value of the compound-free group

The results in Experimental Examples 3-1 to 3-4 are shown in Table 13.

TABLE 13 Estimated tumor Tumor growth Admin- volume inhibitory Exp.istration Dose (test end date) rate Ex. Ex. No. schedule mg/kg/day Mean± S.E. (%) 3-1 0.5% MC qdx17 — 1137 ± 114 Ex. 26 qdx17 25  250 ± 12 78.0qdx17 50   0 ± 0 100 qdx17 100   0 ± 0 100 3-2 0.5% MC qdx18 — 1383 ± 44Ex. 25 qdx18 12.5  926 ± 86 33.0 qdx18 25  250 ± 38 81.9 qdx18 50   0 ±0 100 qdx18 100   0 ± 0 100 Ex. 27 qdx18 25  335 ± 42 75.8 qdx18 50  30± 8 97.8 qdx18 100   0 ± 0 100 3-3 0.5% MC qdx17 —  971 ± 21 Ex. 68qdx17 25 1151 ± 142 0 qdx17 50  559 ± 104 42 qdx17 100  31 ± 23 97 Ex.22 qdx17 25  712 ± 82 27 qdx17 50   9 ± 4 99 qdx17 100   0 ± 0 100 Ex.23 qdx17 25  616 ± 86 37 477 qdx17 50  31 ± 10 97 qdx17 100   0 ± 0 1003-4 0.5% MC qdx16 —  982 ± 103 Ex. 60 qdx16 25  558 ± 79 43 qdx16 50  25± 11 97 qdx16 100   0 ± 0 100 Ex. 67 qdx16 25  896 ± 56 9 qdx16 50  381± 59 61 qdx16 100   0 ± 0 100

[Experimental Example 4] Evaluation of Cell Growth Inhibitory Activity(NPM1 Mutant)

MEM-alpha medium supplemented with 20% FBS was used as a culture mediumfor human AML cells, OCI-AML3 cells. The cells were purchased fromDeutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ).After diluting and preparing each drug (each Example compound shown inTable 14) with Freedom EVO 150 (Tecan Trading AG) (common ratio of 4, 10concentrations from 10 mM to 38 nM), the drug was seeded at 40 nL/wellusing Echo555 (Labcyte Inc.) on a 384-well tissue culture plate (#3712,Corning Inc.) (final concentration 10 gM to 0.038 nM). The prepareddrug-containing plate was stored at −30° C. until use, and thawed beforeuse.

The suspension of the cells was prepared with 10% FBS RPMI1640 culturemedium so as to be 50000 cells/mL, and seeded on the drug-containingplate (40 μL/well) (day 0). The cells were cultured for additional 7days. ATP measurement reagent (CellTiter-Glo (registered trademark,Promega Corporation) 2.0 Assay, model number G9242, Promega Corporation)was added to each well at 10 μL/well, on the day of the drug addition(day 0) and 7 days (day 7) after the drug addition. After affixing ablack sticker on the bottom surface, the mixture was stirred using amicroplate mixing deaeration machine (model name, Weltornado FK-62,Sakaki Dengyo Co., Ltd.) (stirring conditions, revolution 9, rotation 7,time 12). The luminescence (cps) was measured using a luminescencedetector of a microplate reader (model name EnVision 2104-0010,PerkinElmer Co., Ltd.) (N=4).

As an index of the cell growth inhibitory activity, the concentration(GI₅₀) that suppresses cell growth by 50% was calculated using EXCEL2010(Microsoft Corporation). The cell growth of the drug-added group fromday 0 to day 7 was calculated as T/C %, when the cell growth of thedrug-free group was considered as 100%. GI₅o was calculated by GROWTHfunction (exponential regression) using two concentrations sandwichingthe concentration (T/C %=50%) that suppresses cell growth by 50%, andT/C %.

The results in Experimental Example 4 are shown in Table 14.

TABLE 14 Ex. No. GI₅₀ (nM) Day 7 22 217 25 39 27 24 116 46 119 33

[Experimental Example 5] Evaluation of Differentiation-Inducing Activity

The human MLL-AF9 fusion gene was introduced into cKit-positivemononuclear cells isolated from the myeloid of C57BL6 mouse, byretrovirus infection. MLL-AF9-overexpressing AML-like cells (MA9 cells)that acquired abnormal growth ability were established byliquid-culturing the cells after virus infection for a long period oftime. Serum-free medium (GlutaMax, P/S, Stem Pro-34 medium containing 10ng/ml mIL-3, ng/mL mSCF and 10 ng/mL human Oncostatin M) was used as aculture medium.

The suspension of the cells was prepared with the culture medium so asto be 12500 cells/mL, and seeded on a 6-well tissue culture plate at 2mL/well, and then a drug (compound of Example 25, 27, 26 or 22) wasadded thereto at various concentrations (5 nM or 20 nM for the compoundsof Examples 25 and 27, 150 nM or 300 nM for the compound of Example 26,and 50 nM or 100 nM for the compound of Example 22) (2 μL/well) (day 0).For the control group, DMSO was added thereto at a final concentrationof 0.1% (2 μL/well) (day 0). After culturing for 7 days (day 7), thecells were collected, and blocked with 5% FBS/PBS containing 10% MouseBD Fc Block (BD) for 10 minutes at room temperature, and then any ofvarious antibodies (Gr-1-FITC or CD117 (cKit)-APC: Biolegend) was addedthereto at a final concentration of 0.4 ug/sample, and the mixture wasreacted on ice for 30 minutes. Then, dead cell staining dye DAPI (0.2mg/mL) was added thereto at so as to be 1 g L/sample, and the reactionwas carried out on ice for additional 2 minutes under shading. Then, theexpression level of each surface antigen of the cells was measured byNovoCyte flow cytometer (LMS). The obtained data were analyzed by FlowJosoftware (Becton Dickinson), and the rate of each surfaceantigen-expressing cells in the living cells excluding dead cells wasgraphed.

The results in Experimental Example 5 are shown in FIG. 10 and FIG. 11.

FIG. 10 is a graph showing the rate of the myeloid cell differentiationantigen Gr-1-expressing cells in living cells after treatment with thecompound of Example 25, 27, 26 or 22 for 7 days. The vertical axisindicates the percentage of the myeloid cell differentiation antigenGr-1-expressing cells in living cells, and the horizontal axis indicateseach compound and concentration (nM) thereof. Since each compoundincreased the rate of Gr-1-positive cells as compared with the controlgroup, it was demonstrated that these compounds have adifferentiation-inducing action of MA9 cells.

FIG. 11 is a graph showing the rate of the cKit-expressing cells inliving cells after treatment with the compound of Example 25, 27, 26 or22 for 7 days. The vertical axis indicates the percentage of thecKit-expressing cells in living cells, and the horizontal axis indicateseach compound and concentration (nM) thereof. Each compound reduced therate of cKit-positive cells as compared with the control group. cKit isinvolved in the control of survival/differentiation/growth ofhematopoietic progenitor cells, and is particularly highly expressed inimmature hematopoietic stem progenitor cells in myeloid cells. As isclear from the results of FIG. 10 and FIG. 11, it was demonstrated thatthese compounds induce the differentiation of MA9 cells and reduce therate of hematopoietic stem progenitor cells.

[Experimental Example 6] Evaluation of Cell Growth Inhibitory Activityby Combined Use of 2 Drugs

RPMI1640 medium supplemented with 10% FBS was used as a culture mediumfor MOLM-13 cells. The cells were purchased from the American TypeCulture Collection (ATCC). The suspension of MOLM-13 cells was preparedwith 10% FBS RPMI1640 culture medium so as to be 25000 cells/mL, andseeded (50 g L/well) on a 96-well plate (day 0). The solutions of thecompound of Example 25 and any of various other drugs (AraC(Cytarabine), 5Aza (Azacitidine) or Venetoclax) were prepared using aculture solution (the compound of Example 25: common ratio of 4, 5concentrations from the final concentration of 2500 nM, AraC: commonratio of 2, 3 concentrations from the final concentration of 200 nM,5Aza: common ratio of 2, 3 concentrations from the final concentrationof 10000 nM, Venetoclax: common ratio of 2, 3 concentrations from thefinal concentration of 156 nM), and each solution was added to each wellat 25 μL/well, and the mixture was cultured for additional 7 days. Onthe day of the drug addition (day 0) and 7 days after the drug addition(day 7), ATP measurement reagent (CellTiter-Glo (registered trademark,Promega Corporation) 2.0 Assay, model number G9242, Promega Corporation)was added to each well at 50 μL/well. After stirring with a plate mixerfor 2 minutes, the mixture was allowed to stand at room temperature for10 minutes or longer. Then, the luminescence level of each well wasmeasured with a plate reader (N=4, model name EnVision 2104 MultilabelReader, PerkinElmer Co., Ltd.).

The cell growth (%) of the treatment group of the compound of Example 25alone and the treatment group of the compound of Example 25 and theother drug, when the cell growth of the drug-free group was consideredas 100%, from day 0 to day 7, were graphed for each drug using EXCEL2010 (Microsoft Corporation).

FIG. 12 is a graph showing the effects of the combined effect of thecompound of Example 25 and 5Aza on the in-vitro growth of human AML cellline MOLM-13 cells. The vertical axis indicates the cell growth (%), andthe horizontal axis indicates the concentration (nM) of the compound ofExample 25. The symbol black circle indicates the compound of Example 25alone, the symbol black triangle indicates the compound of Example25+5Aza (2.5 μM), the symbol black square indicates the compound ofExample 25+5Aza (5 μM), and the symbol x indicates the compound ofExample 25+5Aza (10 μM). The error bar indicates SD.

FIG. 13 is a graph showing the effects of the combined effect of thecompound of Example 25 and AraC on the in-vitro growth of human AML cellline MOLM-13 cells. The vertical axis indicates the cell growth (%), andthe horizontal axis indicates the concentration (nM) of the compound ofExample 25. The symbol black circle indicates the compound of Example 25alone, the symbol black triangle indicates the compound of Example25+AraC (25 nM), the symbol black square indicates the compound ofExample 25+AraC (50 nM), and the symbol x indicates the compound ofExample 25+AraC (100 nM). The error bar indicates SD.

FIG. 14 is a graph showing the effects of the combined effect of thecompound of Example 25 and Venetoclax on the in-vitro growth of humanAML cell line MOLM-13 cells. The vertical axis indicates the cellgrowth, and the horizontal axis indicates the concentration (nM) of thecompound of Example 25. The symbol black circle indicates the compoundof Example 25 alone, the symbol black triangle indicates the compound ofExample 25+Venetoclax (39 nM), the symbol black square indicates thecompound of Example 25+Venetoclax (78 nM), and the symbol x indicatesthe compound of Example 25+Venetoclax (156 nM). The error bar indicatesSD.

As is clear from these results, it was demonstrated that the combineduse of the compound of Example 25 and any of various drugs enhanced thegrowth inhibitory effect of AML cells.

[Experimental Example 7] Evaluation of Cell Growth Inhibitory Activity

As a culture medium for each cells, RPMI1640 medium supplemented with10% FBS (MOLM-13 cells (human AML cells)) or IMDM medium supplementedwith 10% FBS (K562 cells (human CML cells), MV-4-11 cells (human AMLcells)) was used.

The suspension of each cells was prepared with each culture medium so asto be 25000 cells/mL (K562, MOLM-13) or 50000 cells/mL (MV-4-11), andseeded on a 96-well plate (50 g L/well) (day 0). A solution of eachconcentration (MV-4-11 and MOLM-13: common ratio of 3, 9 concentrationsfrom the final concentration of 1 μM, K562: common ratio of 3, 9concentrations from the final concentration of 10 μM) of a drug (thecompound of Example 131) or a growth medium containing 0.2% DMSO wasadded to each well at 50 μL/well, and the cells were cultured foradditional 7 days. On the day of the drug addition (day 0) and 7 daysafter the drug addition (day 7), ATP measurement reagent (CellTiter-Glo(registered trademark, Promega Corporation) 2.0 Assay, model numberG9242, Promega Corporation) was added to each well at 100 μL/well. Afterstirring with a plate mixer for 2 minutes, the mixture was allowed tostand at room temperature for 10 minutes or longer. Then, theluminescence level of each well was measured with a plate reader (N=6,model name EnVision 2104 Multilabel Reader, PerkinElmer Co., Ltd.).

As an index of the cell growth inhibitory activity, the concentration(GI₅₀) that suppresses cell growth by 50% was calculated using EXCEL2010(Microsoft Corporation). The cell growth of the drug-added group fromday 0 to day 7 was calculated as T/C %, when the cell growth of thedrug-free group (DMSO group) was considered as 100%. The concentration(T/C %=50%) that suppresses cell growth by 50% was calculated as GI₅₀ bySigmoid Emax model. When analysis by the Sigmoid Emax model did notconverge (K562 cells), GI₅₀ was calculated by linear regression usingthe cell viabilities in two concentrations sandwiching 50%. The GI₅₀ ofthe compound of Example 131 in this test was 1.97 nM in MV-4-11 cells,10.1 nM in MOLM-13 cells, and 9110 nM in K562 cells.

INDUSTRIAL APPLICABILITY

Since the compound represented by the general formula (1) or apharmaceutically acceptable salt thereof of the present inventionexhibits an inhibitory action on the interaction between menin and anMLL protein, it can be used for the treatment and/or prophylaxis ofdiseases dependent on the interaction between menin and an MLL protein.Specifically, the compound represented by the general formula (1) or apharmaceutically acceptable salt thereof of the present invention isuseful for the treatment and/or prophylaxis of cancer or diabetes,preferably myelodysplastic syndrome, blood cancer, prostate cancer,breast cancer, hepatoma or pediatric glioma, more preferably leukemia.

1. A compound which is(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olor a pharmaceutically acceptable salt thereof.
 2. The compound accordingto claim 1, which is(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olsuccinate.
 3. The compound according to claim 1, which is represented bythe formula.


4. The compound according to claim 1, which is benzenesulfonate,maleate, fumarate, or hydrochloride of(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-ol.5. A crystal of the compound according to claim 2, wherein the compoundis(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olsuccinate, having at least five peaks at diffraction angles (2θ)selected from 4.66±0.2, 7.02±0.2, 14.10±0.2, 16.68±0.2, 17.46±0.2,18.68±0.2, 21.34±0.2, 24.52±0.2, 25.54±0.2 and 28.22±0.2 in a powderX-ray diffraction diagram obtained through irradiation with copper Kαline (λ=1.54 angstroms).
 6. A crystal of the compound according to claim4, wherein the compound is(1R,2S,4R)-4-({[4-(5,6-dimethoxypyridazin-3-yl)phenyl]methyl}amino)-2-{methyl[6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidin-4-yl]amino}cyclopentan-1-olmaleate, having at least five peaks at diffraction angles (2θ) selectedfrom 4.64±0.2, 7.02±0.2, 7.46±0.2, 11.14±0.2, 14.04±0.2, 16.76±0.2,18.54±0.2, 19.76±0.2, 21.26±0.2 and 22.62±0.2 in a powder X-raydiffraction diagram obtained through irradiation with copper Kα line(λ=1.54 angstroms).
 7. A pharmaceutical composition comprising thecompound according to claim 1, or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.
 8. Thepharmaceutical composition according to claim 7, wherein thepharmaceutically acceptable salt is selected from the group consistingof succinate, benzenesulfonate, maleate, fumarate, and hydrochloride. 9.A pharmaceutical composition comprising the compound according to claim2 and a pharmaceutically acceptable carrier.
 10. A pharmaceuticalcomposition comprising the crystal according to claim 5 and apharmaceutically acceptable carrier.
 11. A pharmaceutical compositioncomprising the compound according to claim 1, or a pharmaceuticallyacceptable salt thereof, one drug selected from the group consisting ofa Bcl-2 inhibitor, a DNA methyltransferase inhibitor, and a pyrimidineantimetabolite, and a pharmaceutically acceptable carrier.
 12. Thepharmaceutical composition according to claim 11, wherein the one drugselected from the group consisting of a Bcl-2 inhibitor, a DNAmethyltransferase inhibitor, and a pyrimidine antimetabolite isVenetoclax.
 13. The pharmaceutical composition according to claim 11,wherein the one drug selected from the group consisting of a Bcl-2inhibitor, a DNA methyltransferase inhibitor, and a pyrimidineantimetabolite is Azacitidine.
 14. The pharmaceutical compositionaccording to claim 11, wherein the one drug selected from the groupconsisting of a Bcl-2 inhibitor, a DNA methyltransferase inhibitor, anda pyrimidine antimetabolite is Cytarabine.
 15. The pharmaceuticalcomposition according to claim 9 further comprising one drug selectedfrom the group consisting of Venetoclax, Azacitidine, and Cytarabine.16. A method for the treatment of acute myelogenous leukemia (AML) oracute lymphocytic leukemia (ALL), the method comprising administering toa subject in need thereof a therapeutically effective amount of thecompound according to claim 1, or a pharmaceutically acceptable saltthereof.
 17. The method according to claim 16, wherein thepharmaceutically acceptable salt is selected from the group consistingof succinate, benzenesulfonate, maleate, fumarate, and hydrochloride.18. The method according to claim 16, wherein the pharmaceuticallyacceptable salt is succinate.
 19. The method according to claim 16,wherein the disease is acute lymphocytic leukemia (ALL).
 20. The methodaccording to claim 16, wherein the disease is acute myelogenous leukemia(AML).
 21. The method according to claim 16, wherein the disease isacute myelogenous leukemia (AML) with NPM1 mutation.
 22. The methodaccording to claim 16 further comprising administering one drug selectedfrom the group consisting of a Bcl-2 inhibitor, a DNA methyltransferaseinhibitor, and a pyrimidine antimetabolite.
 23. A method for thetreatment of acute myelogenous leukemia (AML) or acute lymphocyticleukemia (ALL), the method comprising administering to a subject in needthereof a therapeutically effective amount of the compound according toclaim 1, or a pharmaceutically acceptable salt thereof, and one drugselected from the group consisting of a Bcl-2 inhibitor, a DNAmethyltransferase inhibitor, and a pyrimidine antimetabolite, whereinthe compound, or the pharmaceutically acceptable salt thereof, and theone drug are administered in combination.
 24. A method for the treatmentof acute myelogenous leukemia (AML) or acute lymphocytic leukemia (ALL),the method comprising administering to a subject in need thereof atherapeutically effective amount of the compound according to claim 1,or a pharmaceutically acceptable salt thereof, and one drug selectedfrom the group consisting of a Bcl-2 inhibitor, a DNA methyltransferaseinhibitor and a pyrimidine antimetabolite, wherein the compound, or thepharmaceutically acceptable salt thereof, and the one drug areseparately comprised as active ingredients in different formulations andadministered at the same time or different times.
 25. A method for thetreatment of acute myelogenous leukemia (AML) or acute lymphocyticleukemia (ALL), the method comprising administering to a subject in needthereof a therapeutically effective amount of the compound according toclaim 1, or a pharmaceutically acceptable salt thereof, and one drugselected from the group consisting of a Bcl-2 inhibitor, a DNAmethyltransferase inhibitor and a pyrimidine antimetabolite, wherein thecompound, or the pharmaceutically acceptable salt thereof, and the onedrug are comprised in a single formulation.